TITLE OF THE INVENTION

Antibodies Against Basement Membrane Zone Adhesion Proteins BP 180 and BP230

CROSS-REFERENCE TO RELATED APPLICATION

The present application is entitled to priority under 35 U.S.C. § 119(e) to U.S.

Provisional Patent Application No. 62/741,225 filed October 4, 2018, which is hereby incorporated by reference in its entirety herein.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR

DEVELOPMENT

This invention was made with government support under Grant No. ROl AR052672 awarded by the National Institutes of Health (NIH). The government has certain rights in the invention.

BACKGROUND

Bullous pemphigoid (BP), that mainly affects the elderly, is the most common autoimmune subepidermal blistering skin disease. Patients typically feature tense blisters, but urticarial plaques and pruritus may precede active blistering by weeks to months. Two hemidesmosomal autoantigens were identified as BP 180 (BP antigen, BP AG, 2, collagen XVII) and BP230 (BPAG1, BPAGle), with BP180 being a critical transmembrane protein mediating adhesion of the basal keratinocytes to dermal adhesion proteins such as laminin 332 (Nishie W et al. Am. J Pathol. 2011; 179: 829-37). The non-collagenous domain 16A (NC16A) of the BP180 ectodomain is the immunodominant epitope in BP patients, and antibody titers against BP 180- NC16A correlate to disease activity, as opposed to anti-BP230 antibody titers (Zillikens D, et al. J. Invest. Dermatol. 1997; 109: 573-9; Dopp R, et al. Journal of the American Academy of Dermatology 2000; 42: 577-83; Sitaru C, et al. Experimental dermatology 2007; 16: 770-7; Holtsche MM, et al. The British journal of dermatology 2018; Daneshpazhooh M, et al. Archives of dermatological research 2018; 310: 255-9; Zhou XP, et al. The Journal of dermatology 2016; 43: 141-8; Liu Y, et al. Frontiers in immunology 2017; 8: 1752; Di Zenzo G, et al. The Journal of investigative dermatology 2011; 131 : 2271-80). This finding of a correlation of BP180 but not BP230 antibody titers to disease activity is consistent with an exclusively intracellular expression of BP230 in keratinocytes, making it inaccessible for immunoglobulins, and the immune system, unless keratinocytes are damaged.

Routine diagnostic confirmation and follow-up of BP relies on direct

immunofluorescence (DIF) testing of perilesional biopsies, indirect immunofluorescence (IIF) microscopy of circulating serum antibodies (abs) on monkey esophagus (ME), and/or ELISA testing on recombinantly produced fragments of BP180 (i.e., BP180-NC16A) and BP230 (C-/N- terminal fragments in the MBL system (Hamada T, el al. Experimental dermatology 2001 ; 10: 256-63), C-terminal fragment only in the EUROIMMUN system (Blocker IM, el al. The British journal of dermatology 2012; 166: 964-70)). There are no commercially available test systems for non-NCl6A parts of the BP180 ectodomain and the rod-like middle domain of BP230 (BP230-M).

There is a need in the art for antibodies against basement membrane zone adhesion proteins, such as BP180 and BP230, for methods of making antibodies against basement zone adhesion proteins and methods of their use.

SUMMARY OF THE INVENTION

The present invention provides antibodies against basement membrane zone adhesion proteins BP180 and BP230.

In one aspect, the invention provides a fusion protein comprising a first domain and a second domain, wherein the first domain comprises an antigen-binding domain comprising an antibody or antigen binding fragment thereof that binds BP 180 or BP230, and wherein the second domain comprises a therapeutic polypeptide or fragment thereof.

In certain embodiments, the antibody or antigen binding fragment thereof comprises one CDR selected from the group consisting of at least 1, 2, 3, 4, 5, or 6 CDRs as listed in Table 7.

In certain embodiments, the antibody or antigen binding fragment thereof is a scFv.

In various embodiments of the above aspects or any aspect of the invention delineated herein, the antibody or antigen binding fragment thereof comprises 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 33-64 or 68-70.

In certain embodiments, the therapeutic polypeptide is a complement-inhibiting peptide.

In certain embodiments, the complement-inhibiting peptide inhibits Cls. In certain embodiments, the Cls inhibiting peptide is gigastasin.

In certain embodiments, the complement-inhibiting peptide is a factor H-binding peptide.

In certain embodiments , the factor H-binding peptide is 5C6.

In various embodiments of the above aspects or any aspect of the invention delineated herein, the fusion protein comprises 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 74-76.

In another aspect, the invention provides a recombinant nucleic acid encoding the fusion protein of any one of the above aspects or any aspect of the invention delineated herein.

In another aspect, the invention provides a pharmaceutical composition comprising the fusion protein of any one of the above aspects or any aspect of the invention delineated herein.

In another aspect, the invention provides a method of treating a disease in a subject in need thereof, comprising administering to the subject an effective amount of the pharmaceutical composition of any one of the above aspects or any aspect of the invention delineated herein.

In certain embodiments, the pharmaceutical composition is administered intravenously, intra-articularly, intraperitoneally, intrathecally, intraventricularly, intrasternally, intracranially, intravitreously, intra-muscularly, subcutaneously, orally or intranasally.

In certain embodiments of the above aspects or any aspect of the invention delineated herein, the method further comprises administering a second agent to the subject.

In certain embodiments, the second agent is at least one of carboplatin, cisplatin, paclitaxel, docetaxel, gemcitabine, bevacizumab, olaparib, rucaparib, niraparib,

cyclophosphamide, FU, abiraterone, flutamide, bicalutamide, leuprolide, goserelin, buserelin, triptorelin, degarelix, Enzalutamide, Apalutamide, Sipuleucel-T, Cabazitaxel, Radium-223, trastuzumab, pertuzumab, lapatinib, tamoxifen, oxaliplatin, capecitabine, leucovorin, Irinotecan, Cetuximab, panitumumab, aflibercept, Regorafenib, Trifluridine-tipiracil, immune checkpoint inhibitors (nivolumab, pembrolizumab), cabozantinib, sunitinib, pazopanib, axitinib, interleukin- 2, interferon alpha, mitomycin C, epirubicin, BCG, bleomycin, etoposide, sorafenib, regorafenib, rituximab, dupilumab, efgartigimod, coversin, lenvatinib, pemetrexed and/or vinorelbine.

In another aspect, the invention provides a method of treating a disease in a subject in need thereof, comprising administering to the subject an effective amount of the pharmaceutical composition of any one of the above aspects or any aspect of the invention delineated herein, wherein the fusion protein binds to non-collagenous domain 16A of BP 180, and wherein the disease is bullous pemphigoid, mucous membrane pemphigoid (MMP), linear IgA disease, lichen planus pemphigoides, pemphigoid gestationis, or an autoimmune disease wherein the patient has antibodies against the basement membrane.

In certain embodiments, the fusion protein displaces serum IgG against non-collagenous domain 16A of BP 180, and wherein the complement-inhibiting peptide prevents complement deposits at the BMZ.

In another aspect, the invention provides an isolated antibody or antigen binding fragment thereof that binds BP180 or BP230, wherein the isolated antibody or antigen binding fragment thereof comprises one CDR selected from the group consisting of at least 1, 2, 3, 4, 5 or 6 CDRs as listed in Table 7.

In certain embodiments, the isolated antibody or antigen binding fragment thereof of claim 19, wherein the isolated antibody or antigen binding fragment thereof comprises 91%,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 33-64 or 68-70.

In certain embodiments, the isolated antibody or antigen binding fragment thereof of claim 19, wherein said antibody or antigen binding fragment thereof is non-pathogenic to its target tissue.

In certain embodiments, the isolated antibody or antigen binding fragment cross- competes with anti -BP 180 or anti-BP230 antibodies or fragments thereof.

In certain embodiments, said antibody or antigen binding fragment thereof is human, humanized or chimeric.

In certain embodiments, the isolated antibody or antigen binding fragment thereof of claim 19, wherein said antibody or antigen binding fragment thereof is an scFv.

In another aspect, the invention provides a recombinant nucleic acid encoding the antibody or antigen binding fragment thereof of any one of the above aspects or any aspect of the invention delineated herein.

In another aspect, the invention provides an antibody-drug conjugate comprising the antibody or antigen binding fragment of any one of the above aspects or any other aspect of the invention delineated herein. In certain embodiments of the antibody-drug conjugate, the the drug is MMAE, ozogamicin, emtansine, amanitin, pyrrolobenzodiazepine (PBD) dimer toxin, a chalichaemicin, a cytotoxic maytansinoid, DM1 , carboplatin, cisplatin, paclitaxel, vedotin, or diphtheria toxin.

In another aspect, the invention provides an antibody-protein conjugate. In certain embodiments, the protein is a therapeutic protein. In certain embodiments, the therapeutic protein is an enzyme or a pro-apoptotic protein.

In another aspect, the invention provides a pharmaceutical composition comprising the isolated antibody or antigen binding fragment of any one of the above aspects or any other aspect of the invention delineated herein.

In another aspect, the invention provides a pharmaceutical composition comprising the antibody-drug conjugate of any one of the above aspects or any other aspect of the invention delineated herein.

In another aspect, the invention provides a pharmaceutical composition comprising the antibody-protein conjugate of any one of the above aspects or any other aspect of the invention delineated herein.

In another aspect, the invention provides a method of treating a disease in a subject in need thereof, comprising administering to the subject an effective amount of the pharmaceutical composition of any one of the above aspects or any other aspect of the invention delineated herein.

In certain embodiments, the antibody or antigen binding fragment is administered intravenously, intra-articularly, intraperitoneally, intrathecally, intraventricularly, intrasternally, intracranially, intravitreously, intra-muscularly, subcutaneously, orally or intranasally.

In certain embodiments, the method of treating a disease in a subject in need thereof further comprises administering a second agent to the subject.

In certain embodiments, the second agent is at least one of carboplatin, cisplatin, paclitaxel, docetaxel, gemcitabine, bevacizumab, olaparib, rucaparib, niraparib,

cyclophosphamide, FU, abiraterone, flutamide, bicalutamide, leuprolide, goserelin, buserelin, triptorelin, degarelix, Enzalutamide, Apalutamide, Sipuleucel-T, Cabazitaxel, Radium-223, trastuzumab, pertuzumab, lapatinib, tamoxifen, oxaliplatin, capecitabine, leucovorin, Irinotecan, Cetuximab, panitumumab, aflibercept, Regorafenib, Trifluridine-tipiracil, immune checkpoint inhibitors (nivolumab, pembrolizumab), cabozantinib, sunitinib, pazopanib, axitinib, interleukin- 2, interferon alpha, mitomycin C, epirubicin, BCG, bleomycin, etoposide, sorafenib, regorafenib, rituximab, dupilumab, efgartigimod, coversin, lenvatinib, pemetrexed and/or vinorelbine.

In another aspect, the invention provides a method of treating a disease in a subject in need thereof, comprising administering to the subject an effective amount of the pharmaceutical composition of any of the above aspects or any other aspect of the invention delineated herein, wherein the antibody or antigen binding fragment binds to non-collagenous domain 16A of BP180, and wherein the disease is bullous pemphigoid, mucous membrane pemphigoid (MMP), linear IgA disease, lichen planus pemphigoides, pemphigoid gestationis, or an autoimmune disease wherein the patient has antibodies against the basement membrane.

In certain embodiments, the antibody or antigen binding fragment is an scFv.

In certain embodiments, the antibody or antigen binding fragment displaces serum IgG against non-collagenous domain 16A of BP180.

In another aspect, the invention provides a method of treating a disease in a subject in need thereof, comprising administering to the subject an effective amount of the pharmaceutical composition of any of the above aspects or any aspect of the invention delineated herein, wherein the disease is bullous pemphigoid, mucous membrane pemphigoid (MMP), linear IgA disease, lichen planus pemphigoides, pemphigoid gestationis, anti-p200 pemphigoid, epidermolysis bullosa acquisita, graft versus host disease (GVHD), toxic epidermolysis bullosa, erythema multiforme, other diseases with interface dermatitis, allergic contact dermatitis, atopic dermatitis, psoriasis, vitiligo or skin cancer.

In certain embodiments, the the antibody portion of the antibody-drug conjugate or the antibody-protein conjugate binds to non-collagenous domain 16A of BP180.

In certain embodiments, the antibody or antigen binding fragment is an scFv.

In certain embodiments, the skin cancer is actinic keratosis, squamous cell carcinoma (SCC), basal cell carcinoma (BCC) or melanoma.

BRIEF DESCRIPTION OF THE DRAWINGS

The following detailed description of preferred embodiments of the invention will be better understood when read in conjunction with the appended drawings. For the purpose of illustrating the invention, there are shown in the drawings embodiments which are presently preferred. It should be understood, however, that the invention is not limited to the precise arrangements and instrumentalities of the embodiments shown in the drawings.

Figures 1A-1D illustrate affinity purification (AP) of BP180-NC16A and BP230-C specific antibodies from two representative bullous pemphigoid sera. Eluates from BP180- NCl6A-APs (Figure 1A, Figure 1C) and BP230-C-APs (Figure 1B, Figure 1D) demonstrated monospecific reactivity against the BP230-C or BP180-NC16A antigens, showing good effectivity of the AP procedure.

Figures 2A-2D illustrate indirect immunofluorescence testing of affinity-purified sera on normal human skin and monkey esophagus substrates. By indirect immunofluorescence (IIF) microscopy, affinity-purified BP230-C-specific polyclonal antibodies (abs) showed binding along the basement membrane zone (BMZ) of sections of both normal human skin (NHS)

(Figure 2A) and monkey esophagus (ME) (Figure 2B) substrates; arrowheads point to binding of abs at BMZ. In contrast, BP180-NC16A affinity-purified antibodies stained only on NHS (Figure 2C), but not on ME (Figure 2D) substrate; arrowheads point to binding of abs at BMZ. The observed binding patterns on NHS and ME were confirmed by double IIF with affinity-purified polyclonal IgG and a monoclonal scFv (See Figures 5A-5B).

Figures 3A-3C illustrate that monoclonal antibodies confirm findings from affinity- purified polyclonal antibody testing by indirect immunofluorescence. Antibody phage display- derived scFv mAb clones were confirmed for binding specificity by anti-HA EFISA on BP230-C and BP180-NC16A substrates (Figure 3 A). Deposition of two anti-BP230-C mAbs (BP230-C- mAbl [4-8E, scFv# 28; Table 3] and BP230-C-mAb2 [4-8G, scFv# 14; Table 3]) was observed on NHS and ME substrates (Figure 3B). On the contrary, IIF on ME substrate was completely negative for both anti-NCl6A mAbs tested (BPl80-NCl6A-mAbl [3-4E scFv# 17; Table 3] and BPl80-NCl6A-mAb2 [3-14G, scFv# 1 ; Table 3]) (Figure 3C), but positive for NHS. All mAbs were adjusted to the same concentration of 1 pg/mE before preparing shown dilution series.

Figures 4A-4D illustrate double-depletion experiments of two representative BP sera from BP180-NC16A and BP230-C antibodies, each resulting in positive indirect

immunofluorescence staining on normal human skin (NHS) and monkey esophagus (ME) substrates. Sequential affinity-purification (AP) of two sera on BP180-NC16A, then BP230-C magnetic beads was controlled by standard EFISA before and after AP (Figure 4A, Figure 4C). Indirect immunofluorescence with these eluates resulted in positive linear stainings along the basement membrane zone in both NHS and ME (Figure 4B, Figure 4D), indicating additional serum reactivities against BPl 80-non-NCl6A and BP230-non-C domains/epitopes.

Figures 5A-5B illustrate double indirect immunofluorescence of polyclonal affinity- purified antigen-specific antibodies and a monoclonal antibody. Confirming the binding patterns for anti-BPl 80-NC 16A antibodies seen in Figures 2A-2D and Figures 3A-3C, NC16A expression is present only in normal human skin (Figure 5A), but not in monkey esophagus (Figure 5B), as shown for two representative BP sera (BP-l, BP-2) and the monoclonal antibody BP180-NC16A- m Ab 1.

Figures 6A-6B illustrate that anti-BPl 80-NC 16A XgE variable heavy chain (VH) gene regions feature significantly lower mutation counts when compared to anti -BP 1 80-NC16A IgG VH gene regions, suggesting direct switch from IgM to IgE. Next generation sequencing detects the majority of BP 180-NC 16A-specific IgE clones in the antigen-specific IgGi ₂₃ 4 repertoire of the same patient. Only 2 of 23 anti-BPl 80-NC 16A clones were detected exclusively in IgE (and not IgGi234) repertoire. The majority of BP 180-NC16A IgE mAbs are produced of indirect class- switch recombination, from antigen-specific IgG+ B cells.

Figure 7 illustrates that analysis of light chain usage indicates a predominant utilization of the variable light chain gene VL1-47 in anti-BPl 80-NC16A antibodies obtained by genetic cloning. The same variable light chain gene VL1-47 paired with 12 clonal ly different variable heavy chains in both anti-BPl 80-NC 16 A IgE and IgG Abs, suggesting shared structural and/or ammo acid sequence requirements for binding to the antigen. To confirm findings from genetic clonings, LC-MS/MS was employed on affinity-purified anti-BPl 80-NC 16A serum antibody- light chains from multiple patients (anti-BP230 antibody light chains from same patient sera served as controls). Dominant VL1-47 light chain gene usage may provide a new target for therapy.

Figures 8A-8B are two graphs illustrating the binding of two isolated scFv clones to their target antigen. Various dilutions of 3-14G (Figure 8A) or 3-3E (Figure 8B) were incubated with BP180 protein as a substrate. ELISA was used as a readout. Separate assays using BP230 were used a negative controls.

Figures 9A-9C are a series of micrographs demonstrating the binding of various isolated scFv’s to human skin sections. Normal skin tissue was cryosectioned onto slides and stained with the indicated scFv clones. Indirect immunofluorescence using a FIT C-labe ed, anti-human IgG antibody was then performed to visualize scFv binding. Arrowheads indicate specific staining of the basement membrane.

Figure 10 is a pair of micrographs of an in vitro skin blistering assay demonstrating that anti -BP 180 and anti-BP230 scFVs are non-pathogenic. A mixture of scFv antibodies were incubated with a skin organ culture system prior to staining and assessing for nucroblister formation using indirect immunofluorescence. scFv’s alone do not result in microblister formation, and stain the basement membrane (left panel, arrowheads). Toxicity is only observed when bound scFv’s are further crosshnked using rabbit anti-HA antibody, resulting in microblister formation (right panel, asterisks).

Figures 11 A-l 1 C are a series of graphs demonstrating that anti-BPl 80 scFv antibodies are capable of displacing activity on polyclonal sera from bullous pemphigoid patients.

Polyclonal sera from three BP patients (3419, Figure 11 A; 3389, Figure 1 IB; 3391, Figure 1 1C) was used to coat BP180 substrate in ELISA wells. Increasing concentrations of scFv’s 3-14G (#1), 2-6GL (#29), and 3-3E (#16) were then added to the wells after a brief wash and incubated for 2 hours at 37°C. After two brief washes, new scFv preparations were loaded again to the respective ELISA wells. Displacement of full BP serum IgG antibodies was then detected by developing with anti-human IgG HRP. The anti-BP230 scFv 4-8E.230 (#28) was used as a negative control.

Figures 12A-12E are a series of illustrations, graphs, and micrographs illustrating the construction and characterization of the 1005sqv fusion protein. Figure 12A is a diagram showing the sequence of l005sqv with the various features highlighted (bottom) with a linear listing of the features (top). Figure 12B is a protein gel demonstrating the expression and purification of the lOOSsqv fusion protein. Figure 12C is a reactivity assay demonstrating that 1005sqv is able to specifically bind to BP180 substrate as measured by anti-HA-ELISA. The 3- 14G scFv, which is used as the antigen- binding domain in 1005sqv was used as a positive control. However, 1005sqv w¾s not observed to bind to the basement membrane of human skin sections (Figure 12D), despite being able to inhibit complement fixation by BP antibodies m in vitro complement-fixation assays (Figure 12E).

Figures 13A-13F are a series of illustrations, graphs, and micrographs demonstrating the design and function of the 1012qxm fusion protein. Figure 13 A is a diagram showing a linear description of the features of 1012qxm (top), with a diagram of the sequence (bottom) with the various features highlighted. Figure 13B is a protein gel demonstrating robust production of the fusion protein m eukaryotic cells. 1012qxm was found to possess significant binding specificity for BP180 protein, as measured by ELISA (Figure 13C), as well as by binding the BMZ in human skin sections (Figure 13D) as assessed by indirect immunofluorescence. Staining of BMZ by 1012qxm was likewise found to inhibit complement fixation by BP antibodies, while the 3-14G scFv alone w¾s not (Figure 13E). The reduction in complement fixation was also verified by measuring the concentration of C5a in the reaction supernatants by ELISA (Figure 13F), wliere lG12qxm resulted m significantly lower C5a concentration than 3-14G scFv alone.

Figures 14A-Ί4E are a series of illustrations, graphs, and micrographs demonstrating the design and function of the l048nvm fusion protein. Figure 14A is a diagram showing the linear description of the features of l048nvm (top) with a sequence of the fusion protein with the various features highlighted (bottom). Figure 14B is a protein gel demonstrating robust expression of the fusion protein by eukaryotic cells. Antigen-specificity of 1048nvm w¾s then verified by binding to BP180 protein substrate was measured by anti-HA-ELISA (Figure I4C) Staining of human skin sections with 1048nvm further found robust staining of BMZ, as well as binding to factor H from pooled plasma, demonstrating specific binding of both the anti-BPlBO and complement-specific 5C6 domains (Figure 14D). The ability of l 048nvm to block fixation of C3 as also assessed via indirect immunofluorescence of skin sections stained with 1048nvm followed by incubation with pooled BP serum (Figure 14E).

DETAILED DESCRIPTION

Definitions

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. Although any methods and materials similar or equivalent to those described herein can be used in the practice for testing of the present invention, the preferred materials and methods are described herein. In describing and claiming the present invention, the following terminology will be used.

It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting. The articles“a” and“an” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example,“an element” means one element or more than one element.

“About” as used herein when referring to a measurable value such as an amount, a temporal duration, and the like, is meant to encompass variations of ±20% or ±10%, more preferably ±5%, even more preferably ±1%, and still more preferably ±0.1% from the specified value, as such variations are appropriate to perform the disclosed methods.

As used herein, the term“conservative sequence modifications” is intended to refer to amino acid modifications that do not significantly affect or alter the binding characteristics of the antibody containing the amino acid sequence. Such conservative modifications include amino acid substitutions, additions and deletions. Modifications can be introduced into an antibody of the invention by standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis. Conservative amino acid substitutions are ones in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, one or more amino acid residues within the CDR regions of an antibody can be replaced with other amino acid residues from the same side chain family and the altered antibody can be tested for the ability to bind antigens using the functional assays described herein.

A“disease” is a state of health of an animal wherein the animal cannot maintain homeostasis, and wherein if the disease is not ameliorated then the animal’s health continues to deteriorate. In contrast, a“disorder” in an animal is a state of health in which the animal is able to maintain homeostasis, but in which the animal’s state of health is less favorable than it would be in the absence of the disorder. Left untreated, a disorder does not necessarily cause a further decrease in the animal’s state of health. In some embodiments, the animal is a human.

“Effective amount” or“therapeutically effective amount” are used interchangeably herein, and refer to an amount of a compound, formulation, material, or composition, as described herein effective to achieve a particular biological result or provides a therapeutic or prophylactic benefit. Such results may include, but are not limited to, anti-tumor activity as determined by any means suitable in the art.

“Encoding” refers to the inherent property of specific sequences of nucleotides in a polynucleotide, such as a gene, a cDNA, or an mRNA, to serve as templates for synthesis of other polymers and macromolecules in biological processes having either a defined sequence of nucleotides (i.e., rRNA, tRNA and mRNA) or a defined sequence of amino acids and the biological properties resulting therefrom. Thus, a gene encodes a protein if transcription and translation of mRNA corresponding to that gene produces the protein in a cell or other biological system. Both the coding strand, the nucleotide sequence of which is identical to the mRNA sequence and is usually provided in sequence listings, and the non-coding strand, used as the template for transcription of a gene or cDNA, can be referred to as encoding the protein or other product of that gene or cDNA.

As used herein“endogenous” refers to any material from or produced inside an organism, cell, tissue or system.

As used herein, the term“exogenous” refers to any material introduced from or produced outside an organism, cell, tissue or system.

The term“expression” as used herein is defined as the transcription and/or translation of a particular nucleotide sequence driven by its promoter.

“Expression vector” refers to a vector comprising a recombinant polynucleotide comprising expression control sequences operatively linked to a nucleotide sequence to be expressed. An expression vector comprises sufficient cis-acting elements for expression; other elements for expression can be supplied by the host cell or in an in vitro expression system. Expression vectors include all those known in the art, such as cosmids, plasmids ( e.g ., naked or contained in liposomes) and viruses (e.g., sendai viruses, lentiviruses, retroviruses, adenoviruses, and adeno-associated viruses) that incorporate the recombinant polynucleotide.

“Homologous” as used herein, refers to the subunit sequence identity between two polymeric molecules, e.g., between two nucleic acid molecules, such as, two DNA molecules or two RNA molecules, or between two polypeptide molecules. When a subunit position in both of the two molecules is occupied by the same monomeric subunit; e.g., if a position in each of two DNA molecules is occupied by adenine, then they are homologous at that position. The homology between two sequences is a direct function of the number of matching or homologous positions; e.g., if half (e.g., five positions in a polymer ten subunits in length) of the positions in two sequences are homologous, the two sequences are 50% homologous; if 90% of the positions (e.g., 9 of 10), are matched or homologous, the two sequences are 90% homologous.

“Humanized” forms of non-human (e.g., murine) antibodies are chimeric

immunoglobulins, immunoglobulin chains or fragments thereof (such as Fv, Fab, Fab', F(ab')2 or other antigen-binding subsequences of antibodies) which contain minimal sequence derived from non-human immunoglobulin. For the most part, humanized antibodies are human

immunoglobulins (recipient antibody) in which residues from a complementary-determining region (CDR) of the recipient are replaced by residues from a CDR of a non-human species (donor antibody) such as mouse, rat or rabbit having the desired specificity, affinity, and capacity. In some instances, Fv framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies can comprise residues which are found neither in the recipient antibody nor in the imported CDR or framework sequences. These modifications are made to further refine and optimize antibody performance. In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin sequence. The humanized antibody optimally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see Jones et al, Nature, 321 : 522-525, 1986;

Reichmann et al., Nature, 332: 323-329, 1988; Presta, Curr. Op. Struct. Biol., 2: 593-596, 1992.

“Fully human” refers to an immunoglobulin, such as an antibody, where the whole molecule is of human origin or consists of an amino acid sequence identical to a human form of the antibody.

“Identity” as used herein refers to the subunit sequence identity between two polymeric molecules particularly between two amino acid molecules, such as, between two polypeptide molecules. When two amino acid sequences have the same residues at the same positions; e.g., if a position in each of two polypeptide molecules is occupied by an Arginine, then they are identical at that position. The identity or extent to which two amino acid sequences have the same residues at the same positions in an alignment is often expressed as a percentage. The identity between two amino acid sequences is a direct function of the number of matching or identical positions; e.g., if half (e.g., five positions in a polymer ten amino acids in length) of the positions in two sequences are identical, the two sequences are 50% identical; if 90% of the positions (e.g., 9 of 10), are matched or identical, the two amino acids sequences are 90% identical.

The term“immune response” as used herein is defined as a cellular response to an antigen that occurs when lymphocytes identify antigenic molecules as foreign and induce the formation of antibodies and/or activate lymphocytes to remove the antigen.

As used herein, an“instructional material” includes a publication, a recording, a diagram, or any other medium of expression which can be used to communicate the usefulness of the compositions and methods of the invention. The instructional material of the kit of the invention may, for example, be affixed to a container which contains the nucleic acid, peptide, and/or composition of the invention or be shipped together with a container which contains the nucleic acid, peptide, and/or composition. Alternatively, the instructional material may be shipped separately from the container with the intention that the instructional material and the compound be used cooperatively by the recipient.

“Isolated” means altered or removed from the natural state. For example, a nucleic acid or a peptide naturally present in a living animal is not“isolated,” but the same nucleic acid or peptide partially or completely separated from the coexisting materials of its natural state is “isolated.” An isolated nucleic acid or protein can exist in substantially purified form, or can exist in a non-native environment such as, for example, a host cell.

A“lentivirus” as used herein refers to a genus of the Retroviridae family. Lentiviruses are unique among the retroviruses in being able to infect non-dividing cells; they can deliver a significant amount of genetic information into the DNA of the host cell, so they are one of the most efficient methods of a gene delivery vector. HIV, SIV, and FIV are all examples of lentiviruses. Vectors derived from lentiviruses offer the means to achieve significant levels of gene transfer in vivo.

By the term“modified” as used herein, is meant a changed state or structure of a molecule or cell of the invention. Molecules may be modified in many ways, including chemically, structurally, and functionally. Cells may be modified through the introduction of nucleic acids. By the term“modulating,” as used herein, is meant mediating a detectable increase or decrease in the level of a response in a subject compared with the level of a response in the subject in the absence of a treatment or compound, and/or compared with the level of a response in an otherwise identical but untreated subject. The term encompasses perturbing and/or affecting a native signal or response thereby mediating a beneficial therapeutic response in a subject, preferably, a human.

In the context of the present invention, the following abbreviations for the commonly occurring nucleic acid bases are used. “A” refers to adenosine,“C” refers to cytosine,“G” refers to guanosine,“T” refers to thymidine, and“U” refers to uridine.

Unless otherwise specified, a“nucleotide sequence encoding an amino acid sequence” includes all nucleotide sequences that are degenerate versions of each other and that encode the same amino acid sequence. The phrase nucleotide sequence that encodes a protein or an RNA may also include introns to the extent that the nucleotide sequence encoding the protein may in some version contain an intron(s).

The term“operably linked” refers to functional linkage between a regulatory sequence and a heterologous nucleic acid sequence resulting in expression of the latter. For example, a first nucleic acid sequence is operably linked with a second nucleic acid sequence when the first nucleic acid sequence is placed in a functional relationship with the second nucleic acid sequence. For instance, a promoter is operably linked to a coding sequence if the promoter affects the transcription or expression of the coding sequence. Generally, operably linked DNA sequences are contiguous and, where necessary to join two protein coding regions, in the same reading frame.

The term“overexpressed” tumor antigen or“overexpression” of a tumor antigen is intended to indicate an abnormal level of expression of a tumor antigen in a cell from a disease area like a solid tumor within a specific tissue or organ of the patient relative to the level of expression in a normal cell from that tissue or organ. Patients having solid tumors or a hematological malignancy characterized by overexpression of the tumor antigen can be determined by standard assays known in the art.

“Parenteral” administration of an immunogenic composition includes, e.g., subcutaneous (s.c.), intravenous (i.v.), intramuscular (i.m.), or intrasternal injection, or infusion techniques. The term“polynucleotide” as used herein is defined as a chain of nucleotides.

Furthermore, nucleic acids are polymers of nucleotides. Thus, nucleic acids and polynucleotides as used herein are interchangeable. One skilled in the art has the general knowledge that nucleic acids are polynucleotides, which can be hydrolyzed into the monomeric“nucleotides.” The monomeric nucleotides can be hydrolyzed into nucleosides. As used herein polynucleotides include, but are not limited to, all nucleic acid sequences which are obtained by any means available in the art, including, without limitation, recombinant means, i.e., the cloning of nucleic acid sequences from a recombinant library or a cell genome, using ordinary cloning technology and PCR™, and the like, and by synthetic means.

As used herein, the terms“peptide,”“polypeptide,” and“protein” are used

interchangeably, and refer to a compound comprised of amino acid residues covalently linked by peptide bonds. A protein or peptide must contain at least two amino acids, and no limitation is placed on the maximum number of amino acids that can comprise a protein’s or peptide’s sequence. Polypeptides include any peptide or protein comprising two or more amino acids joined to each other by peptide bonds. As used herein, the term refers to both short chains, which also commonly are referred to in the art as peptides, oligopeptides and oligomers, for example, and to longer chains, which generally are referred to in the art as proteins, of which there are many types. “Polypeptides” include, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, fusion proteins, among others. The polypeptides include natural peptides, recombinant peptides, synthetic peptides, or a combination thereof.

The term“promoter” as used herein is defined as a DNA sequence recognized by the synthetic machinery of the cell, or introduced synthetic machinery, required to initiate the specific transcription of a polynucleotide sequence.

As used herein, the term“promoter/regulatory sequence” means a nucleic acid sequence which is required for expression of a gene product operably linked to the promoter/regulatory sequence. In some instances, this sequence may be the core promoter sequence and in other instances, this sequence may also include an enhancer sequence and other regulatory elements which are required for expression of the gene product. The promoter/regulatory sequence may, for example, be one which expresses the gene product in a tissue specific manner. A“constitutive” promoter is a nucleotide sequence which, when operably linked with a polynucleotide which encodes or specifies a gene product, causes the gene product to be produced in a cell under most or all physiological conditions of the cell.

An“inducible” promoter is a nucleotide sequence which, when operably linked with a polynucleotide which encodes or specifies a gene product, causes the gene product to be produced in a cell substantially only when an inducer which corresponds to the promoter is present in the cell.

A“tissue-specific” promoter is a nucleotide sequence which, when operably linked with a polynucleotide encodes or specified by a gene, causes the gene product to be produced in a cell substantially only if the cell is a cell of the tissue type corresponding to the promoter.

A“Sendai virus” refers to a genus of the Paramyxoviridae family. Sendai viruses are negative, single stranded RNA viruses that do not integrate into the host genome or alter the genetic information of the host cell. Sendai viruses have an exceptionally broad host range and are not pathogenic to humans. Used as a recombinant viral vector, Sendai viruses are capable of transient but strong gene expression

A“signal transduction pathway” refers to the biochemical relationship between a variety of signal transduction molecules that play a role in the transmission of a signal from one portion of a cell to another portion of a cell. The phrase“cell surface receptor” includes molecules and complexes of molecules capable of receiving a signal and transmitting signal across the plasma membrane of a cell.

“Single chain antibodies” refer to antibodies formed by recombinant DNA techniques in which immunoglobulin heavy and light chain fragments are linked to the Fv region via an engineered span of amino acids. Various methods of generating single chain antibodies are known, including those described in U.S. Pat. No. 4,694,778; Bird (1988) Science 242:423-442; Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883; Ward et al. (1989) Nature 334:54454; Skerra et al. (1988) Science 242: 1038-1041.

By the term“specifically binds,” as used herein with respect to an antibody, is meant an antibody which recognizes a specific antigen, but does not substantially recognize or bind other molecules in a sample. For example, an antibody that specifically binds to an antigen from one species may also bind to that antigen from one or more species. But, such cross-species reactivity does not itself alter the classification of an antibody as specific. In another example, an antibody that specifically binds to an antigen may also bind to different allelic forms of the antigen.

However, such cross reactivity does not itself alter the classification of an antibody as specific.

In some instances, the terms“specific binding” or“specifically binding,” can be used in reference to the interaction of an antibody, a protein, or a peptide with a second chemical species, to mean that the interaction is dependent upon the presence of a particular structure (e.g., an antigenic determinant or epitope) on the chemical species; for example, an antibody recognizes and binds to a specific protein structure rather than to proteins generally. If an antibody is specific for epitope“A”, the presence of a molecule containing epitope A (or free, unlabeled A), in a reaction containing labeled“A” and the antibody, will reduce the amount of labeled A bound to the antibody.

The term“subject” is intended to include living organisms in which an immune response can be elicited (e.g., mammals). A“subject” or“patient,” as used therein, may be a human or non-human mammal. Non-human mammals include, for example, livestock and pets, such as ovine, bovine, porcine, canine, feline and murine mammals. Preferably, the subject is human.

As used herein, a“substantially purified” cell is a cell that is essentially free of other cell types. A substantially purified cell also refers to a cell which has been separated from other cell types with which it is normally associated in its naturally occurring state. In some instances, a population of substantially purified cells refers to a homogenous population of cells. In other instances, this term refers simply to cell that have been separated from the cells with which they are naturally associated in their natural state. In some embodiments, the cells are cultured in vitro. In other embodiments, the cells are not cultured in vitro.

The term“therapeutic” as used herein means a treatment and/or prophylaxis. A therapeutic effect is obtained by suppression, remission, or eradication of a disease state.

The term“transfected” or“transformed” or“transduced” as used herein refers to a process by which exogenous nucleic acid is transferred or introduced into the host cell. A “transfected” or“transformed” or“transduced” cell is one which has been transfected, transformed or transduced with exogenous nucleic acid. The cell includes the primary subject cell and its progeny.

To“treat” a disease as the term is used herein, means to reduce the frequency or severity of at least one sign or symptom of a disease or disorder experienced by a subject. The term“antibody,” as used herein, refers to an immunoglobulin molecule which is able to specifically bind to a specific epitope on an antigen.

Antibodies can be intact immunoglobulins derived from natural sources or from recombinant sources and can be immunoreactive portions of intact immunoglobulins. Some antibodies are typically tetramers of immunoglobulin molecules. The antibodies in the present invention may exist in a variety of forms including, for example, polyclonal antibodies, monoclonal antibodies, chimeric, hybrid, Fv, Fab, F(ab) ₂ , scFv, single chain antibodies, primatized and humanized antibodies. Antibody fragments refer to antigen-binding

immunoglobulin peptides which are at least about 5 to about 15 amino acids or more in length, and which retain the capacity to bind to the antigen. (Harlow et al, 1999, Using Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, NY; Harlow et al, 1989, Antibodies: A Laboratory Manual, Cold Spring Harbor, N.Y.; Houston et al, 1988, Proc. Natl. Acad. Sci. USA 85:5879-5883; Bird et al., 1988, Science 242:423-426).

An“antibody heavy chain,” as used herein, refers to the larger of the two types of polypeptide chains present in all antibody molecules in their naturally occurring conformations.

An“antibody light chain,” as used herein, refers to the smaller of the two types of polypeptide chains present in all antibody molecules in their naturally occurring conformations.

By the term“synthetic antibody or antigen binding fragment thereof’ as used herein, is meant an antibody which is generated using recombinant DNA technology, such as, for example, an antibody expressed by a bacteriophage as described herein. The term should also be construed to mean an antibody or antigen binding fragment thereof which has been generated by the synthesis of a DNA molecule encoding the antibody or antigen binding fragment thereof and which DNA molecule expresses an antibody or antigen binding fragment thereof protein, or an amino acid sequence specifying the antibody or antigen binding fragment thereof, wherein the DNA or amino acid sequence has been obtained using synthetic DNA or amino acid sequence technology which is available and well known in the art.

By the term“Fab/phage” as used herein, is meant a phage particle which expresses the Fab portion of an antibody.

The term“antibody-protein fusion molecule” as used herein, refers to a fusion or chimeric molecule comprising at least two components, including a targeting moiety and peptide or protein. The fusion molecule can also comprise at least two components, including a targeting moiety and a therapeutic moiety. In other embodiments, the fusion molecule can comprise at least two components, including a targeting moiety and a non-protein molecule.

As used herein,“non-pathogenic” when used in conjuction with the term“antibody” refers to an antibody that does not appreciably alter normal homeostasis of a mammal when introduced into the mammal. Preferably, the non-pathogenic antibody does not have an appreciable adverse effect on the host mammal when present in the host mammal.

By the term“scFv/phage” as used herein, is meant a phage particle which expresses the Fv portion of an antibody as a single chain.

“Phage,” or“phage particle,” as these terms are used herein, inclue particles that contain phage nuclei encoding, inter alia, an antibody. This is because, as would be appreciated by the skilled artisan, unlike peptide phage display (where the peptide DNA insert is small and it is actually cloned into the phage DNA), the larger scFv or Fab DNA inserts are actually cloned into, among other things, a plasmid. Thus, the nucleic acid encoding the antibody, e.g., a plasmid such as, but not limited to, pComb3X, not only comprises a plasmid origin of replication, but also a phage (e.g. Ml 3) origin of replication sequence and an Ml 3 packaging sequence, so that when the nucleic acid is produced, a helper phage can be used to provide the required phage (e.g., Ml 3) proteins in trans to make“phage-like” particles. That is, these particles resemble phage on the outside, but on the inside they contain plasmid (also referred to as“phagemid”) DNA. In other words, the phagemid DNA need not encode any Ml 3 phage proteins, except a piece of Ml 3 gene III fused to the DNA for antibody or peptide. Thus, it should be understood that the terms“phage,”“phage particle,”“phage-like particle” and“phagemid” are used interchangeably herein.

As used herein, the term“washing” refers to removing at least one component from a mixture of at least two components. By way of a series of non-limiting examples, salt can be washed from a protein by dialyzing a protein, an antibody can be removed from the outside of a ce4ll by altering the salt conditions of the cell medium or by removing the salt from the cell medium altogether, and an unbound phage can be removed from a cell suspension by separating the cell from the phage using a gel filtration technique.

As used herein, the terms“therapeutic protein” and“therapeutic polypeptide” refer to a protein, a polypeptide, an antibody, a peptide or fragment or variant thereof, having one or more therapeutic and/or biological activity. Therapeutic proteins and polypeptides encompassed by the invention include but are not limited to, proteins, polypeptides, peptides, antibodies, and biologies. The terms peptides, proteins, and polypeptides are used interchangeably herein. Thus, a fusion protein of the invention may contain at least a fragment or variant of a therapeutic protein, and/or at least a fragment or variant of an antibody. Additionally, the term“therapeutic protein” may refer to the endogenous or naturally occurring correlate of a therapeutic protein.

By a polypeptide displaying a“therapeutic activity” or a protein that is“therapeutically active” is meant a polypeptide that possesses one or more known biological and/ or therapeutic activities associated with a therapeutic protein such as one or more of the therapeutic proteins described herein or otherwise known in the art. As a non-limiting example, a“therapeutic protein” is a protein that is useful to treat, prevent or ameliorate a disease, condition or disorder. For example, a non-exhaustive list of“therapeutic protein” portions which may be comprised by a fusion protein of the invention includes, but is not limited to Fas ligand, tumor necrosis factor alpha receptor, CD200, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), transforming growth factor alpha (TGF alpha), programmed death ligand (PD-L1), and epidermal growth factor. Other useful therapeutic proteins include enzymes, such as those that activate drugs, or those that can act to inhibit an undesirable biological reaction in a mammal. Likewise, other useful therapeutic proteins can bind and thus inhibit undesirable or disease- related biological reactions in mammals.

A "targeting moiety" or a "cell-specific targeting moiety" is used herein to refer to a molecule comprising an antibody to one or more particular soluble protein, tissue marker, cell surface antigen, cell marker, growth factor, hormone, or cytokine. In one aspect, the targeting moiety of a chimeric composition is an antibody, such as a single chain antibody, an antibody fragment, a Fab, and the likes.

As used herein, "pro-apoptotic molecules" refers to any molecule that is capable of inducing cell death through apoptotic mechanisms. The pro-apoptotic molecules can induce programmed cell death upon entry into the target cell. Apoptosis, or programmed cell death, is a fundamental process controlling normal tissue homeostasis by regulating a balance between cell proliferation and death. Examples of pro-apoptotic molecules include but are not limited to granzymes, Bcl-2 family members (Bax, Bak, Bcl-Xs, Bik, Bok, Bipla, and the like), and caspases. By "laser absorption molecule," as the term is used herein, is meant a molecule that can absorb laser light radiation. The term is also used to indicate a molecule that can absorb any type of laser energy, including light-based laser energy, but also including any form of electromagnetic or particle-based laser energy.

As used herein, the term "laser absorbing molecule complex" refers to a complex comprising at least one laser absorbing molecule and at least one antibody or scFv of the invention. A complex can have the two or molecules associated by way of a non-covalent interaction, or by way of one or more covalent interactions.

The phrase“under transcriptional control” or“operatively linked” as used herein means that the promoter is in the correct location and orientation in relation to a polynucleotide to control the initiation of transcription by RNA polymerase and expression of the polynucleotide.

A“vector” is a composition of matter which comprises an isolated nucleic acid and which can be used to deliver the isolated nucleic acid to the interior of a cell. Numerous vectors are known in the art including, but not limited to, linear polynucleotides, polynucleotides associated with ionic or amphiphilic compounds, plasmids, and viruses. Thus, the term“vector” includes an autonomously replicating plasmid or a virus. The term should also be construed to include non-plasmid and non-viral compounds which facilitate transfer of nucleic acid into cells, such as, for example, polylysine compounds, liposomes, and the like. Examples of viral vectors include, but are not limited to, Sendai viral vectors, adenoviral vectors, adeno-associated virus vectors, retroviral vectors, lentiviral vectors, and the like.

As used herein, the term "genetic construct" refers to the DNA or RNA molecules that comprise a nucleotide sequence which encodes protein. The coding sequence includes initiation and termination signals operably linked to regulatory elements including a promoter and polyadenylation signal capable of directing expression in the cells of the individual to whom the nucleic acid molecule is administered.

Ranges: throughout this disclosure, various aspects of the invention can be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range. Description

The present invention provides a fusion protein comprising two domains. One of the domains is an antigen-binding domain comprising an antibody or antigen binding fragment thereof that binds the human basement membrane zone proteins BP180 or BP230. The second domain comprises a therapeutic polypeptide or fragment thereof. In some embodiments, the antibody or antigen binding fragment thereof of the antigen-binding domain comprises one CDR selected from the group consisting of at least 1, 2, 3, 4, 5, or 6 CDRs as listed in Table 7. In further embodiments, the antibody or antigen binding fragment thereof of the antigen-binding domain is a scFv. In some embodiments of the fusion protein, the antibody or antigen binding fragment thereof comprises 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 33-64 or 68-70.

In some embodiments of the fusion protein, the therapeutic polypeptide or fragment thereof inhibits components of the complement system. In some embodiments, the

complement-inhibiting polypeptide binds and inhibits complement component Cls. By way of a non-limiting example, the Cls inhibiting polypeptide is gigastasin. In some embodiments, the complement-inhibiting polypeptide binds and inhibits the complement-regulating factor H. By way of a non-limiting example, the factor H inhibitor is 5C6 peptide. In some embodiments, the fusion protein of the invention comprises 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 74-76.

Accordingly, the invention further provides a recombinant nucleic acid encoding the fusion protein.

Provided is a pharmaceutical composition comprising the fusion protein of any one of the previous embodiments, as well as a method of treating a disease in a subject in need thereof, comprising administering to the subject an effective amount of the pharmaceutical composition comprising the fusion protein of any one of the previous embodiments. The pharmaceutical composition is administered intravenously, intra-articularly, intraperitoneally, intrathecally, intraventricularly, intrasternally, intracranially, intravitreously, intra-muscularly, subcutaneously, orally or intranasally. In some embodiments, composition of the invention is further administerd with a second agent to the subject. In some embodiments, the second agent is at least one of carboplatin, cisplatin, paclitaxel, docetaxel, gemcitabine, bevacizumab, olaparib, rucaparib, niraparib, cyclophosphamide, FU, abiraterone, flutamide, bicalutamide, leuprolide, goserelin, buserelin, triptorelin, degarelix, Enzalutamide, Apalutamide, Sipuleucel-T, Cabazitaxel, Radium- 223, trastuzumab, pertuzumab, lapatinib, tamoxifen, oxaliplatin, capecitabine, leucovorin, Irinotecan, Cetuximab, panitumumab, aflibercept, Regorafenib, Trifluridine-tipiracil, immune checkpoint inhibitors (nivolumab, pembrolizumab), cabozantinib, sunitinib, pazopanib, axitinib, interleukin-2, interferon alpha, mitomycin C, epirubicin, BCG, bleomycin, etoposide, sorafenib, regorafenib, rituximab, dupilumab, efgartigimod, coversin, lenvatinib, pemetrexed and/or vinorelbine.

Also provided is a method of treating a disease in a subject in need thereof, comprising administering to the subject an effective amount of the pharmaceutical composition of any one of the previous embodiments, wherein the fusion protein binds to non-collagenous domain 16A of the BP 180 protein in the human basement membrane zone, and wherein the disease is bullous pemphigoid, mucous membrane pemphigoid (MMP), linear IgA disease, lichen planus pemphigoides, pemphigoid gestationis, or an autoimmune disease wherein the patient has auto antibodies against the basement membrane.

One skilled in the art armed with the present specification would understand that in some embodiments, the antigen-binding domain of the fusion protein of any one of the previous embodiments displaces auto-reactive, disease-causing serum IgG against non-collagenous domain 16A of BP 180, and the complement-inhibiting therapeutic peptide prevents complement deposits at the BMZ, thereby achieving treatment of the disease.

Antibodies

Provided is an isolated antibody or antigen binding fragment thereof that binds BP180 or BP230, wherein the isolated antibody or antigen binding fragment thereof comprises at least 1, 2, 3, 4, 5 or 6 CDRs from Table 7. In some embodiments, the CDRs are from the same antibody in

Table 7. In some embodiments, the CDRs are mixed from different antibodies in Table 7 that bind to the same protein (BP180 or BP230). In some embodiments, the antibody or antigen binding fragment thereof binds BP 180. In further embodiments, the antibody or antigen binding fragment thereof binds the non-collagenous domain 16A (NC16A) of the BP180 ectodomain. In some embodiments, the antibody or antigen binding fragment thereof binds BP230.

In some embodiments, the isolated antibody or antigen binding fragment thereof comprises 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, and any partial integers therebetween, sequence identity to the amino acid sequence of any one of SEQ ID NOs: 33-64 or 68-70.

In some embodiments, the antibody or antigen binding fragment thereof is non- pathogenic to its target tissue.

In some embodiments, the antibody or antigen binding fragment thereof cross-competes with anti-BPl80 or anti-BP230 antibodies or fragments thereof. In further embodiments, the antibody or antigen binding fragment thereof cross-competes with anti-BPl 80-NCl6A antibodies or fragments thereof.

In some embodiments, the antibody or antigen binding fragment thereof is humanized or chimeric.

In some embodiments, the antibody or antigen binding fragment thereof is an scFv.

Also provided is a recombinant nucleic acid encoding the antibody or antigen binding fragment thereof of any one of the previous embodiments.

Also provided is an antibody-drug conjugate comprising the antibody or antigen binding fragment of any one of the previous embodiments. In some embodiments, the drug is MMAE, ozogamicin, emtansine, amanitin, pyrrolobenzodiazepine (PBD) dimer toxin, a chalichaemicin, a cytotoxic maytansinoid, DM1 , carboplatin, cisplatin, paclitaxel, vedotin, or diphtheria toxin.

Also provided is an antibody-protein conjugate comprising the antibody or antigen binding fragment of any one of the previous embodiments. In some embodiments, the protein is a therapeutic protein. In some embodiments, the therapeutic protein is an enzyme or a pro- apoptotic protein. In some embodiments, the protein is an inhibitor of complement cascade, for example inhibiting the classical pathway and/or the alternative pathway and/or the lectin pathway. In further embodiments, the protein is a cytokine, for example IL-2. In some embodiments, the protein is a cytokine-receptor inhibitor such as dupilumab. In some embodiments, the protein is a cell surface protein. In further embodiments, the protein is PD-l. Provided is a pharmaceutical composition comprising the isolated antibody or antigen binding fragment, or the antibody-drug conjugate or the antibody-protein conjugate of any one of the previous embodiments.

Provided is a method of treating a disease in a subject in need thereof, comprising administering to the subject an effective amount of the pharmaceutical composition of any one of the previous embodiments. In some embodiments, the antibody or antigen binding fragment is administered intravenously, intra-articularly, intraperitoneally, intrathecally, intraventricularly, intrasternally, intracranially, intra-muscularly, intra-vitreously, subcutaneously, orally or intranasally. In some embodiments, the method further comprises administering a second agent to the subject. In further embodiments, the second agent is at least one of carboplatin, cisplatin, paclitaxel, docetaxel, gemcitabine, bevacizumab, olaparib, rucaparib, niraparib,

cyclophosphamide, FU, abiraterone, flutamide, bicalutamide, leuprobde, goserebn, buserebn, triptorebn, degarelix, Enzalutamide, Apalutamide, Sipuleucel-T, Cabazitaxel, Radium-223, trastuzumab, pertuzumab, lapatinib, tamoxifen, oxaliplatin, capecitabine, leucovorin, Irinotecan, Cetuximab, panitumumab, aflibercept, Regorafenib, Trifluridine-tipiracil, immune checkpoint inhibitors (nivolumab, pembrolizumab), cabozantinib, sunitinib, pazopanib, axitinib, interleukin- 2, interferon alpha, mitomycin C, epirubicin, BCG, bleomycin, etoposide, sorafenib, regorafenib, rituximab, dupilumab, efgartigimod, coversin, lenvatinib, pemetrexed and/or vinorelbine. In some embodiments, the second agent is an alkylating agent, antimetabobte, antibiotic, a plant- derived agent, platinum complex, campthotecin derivative, tyrosine kinase inhibitor, monoclonal antibody, interferon, biological response modifier, hormonal anti-tumor agent, anti-tumor viral agent, angiogenesis inhibitor, differentiating agent, PI3K/mTOR/AKT inhibitor, cell cycle inhibitor, apoptosis inhibitor, hsp 90 inhibitor, tubulin inhibitor, DNA repair inhibitor, anti- angiogenic agent, receptor tyrosine kinase inhibitor, topoisomerase inhibitor, taxane, agent targeting Her-2, hormone antagonist, agent targeting a growth factor receptor, or a

pharmaceutically acceptable salt thereof. In some embodiments, the anti-tumor agent is citabine, capecitabine, valopicitabine or gemcitabine. In some embodiments, the anti-tumor agent is Avastin, Sutent, Nexavar, Recentin, ABT-869, Axitinib, Irinotecan, topotecan, paclitaxel, docetaxel, lapatinib, Herceptin, tamoxifen, progesterone, a steroidal aromatase inhibitor, a non steroidal aromatase inhibitor, Fulvestrant, an inhibitor of epidermal growth factor receptor (EGFR), Cetuximab, Panitumimab, an inhibitor of insulin-like growth factor 1 receptor (IGF1R), and/or CP-751871.

Provided is a method of treating a disease in a subject in need thereof, comprising administering to the subject an effective amount of a pharmaceutical composition comprising the isolated antibody or antibody fragment of any one of the previous embodiments. In some embodiments, the antibody or antibody fragment binds to non-collagenous domain 16A of BP 180. In some embodiments, the disease is bullous pemphigoid, mucous membrane

pemphigoid (MMP), linear IgA disease, lichen planus pemphigoides or pemphigoid gestationis. In some embodiments, the antibody or antigen binding fragment is an scFv. In further embodiments, the antibody or antigen binding fragment displaces serum IgG against non- collagenous domain 16A of BP 180.

Provided is a method of treating a disease in a subject in need thereof, comprising administering to the subject an effective amount of a pharmaceutical composition comprising the antibody-protein conjugate or the antibody-drug conjugate of any one of the previous

embodiments. In some embodiments, the antibody or antibody fragment binds to non- collagenous domain 16A of BP180. In further embodiments, the antibody or antibody fragment that binds to non-collagenous domain 16A of BP180 is listed in Table 3. In yet further embodiments, the antibody or antigen binding fragment thereof comprises 91%, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99% or 100%, and any partial integers therebetween, sequence identity to the amino acid sequence of any one of SEQ ID NOs: 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 61, 62, 63, 64, and 70. In some

embodiments, the disease is bullous pemphigoid, mucous membrane pemphigoid (MMP), linear IgA disease, lichen planus pemphigoides, pemphigoid gestationis, anti-p200 pemphigoid, epidermolysis bullosa acquisita, graft versus host disease (GVHD), toxic epidermolysis bullosa, erythema multiforme, other diseases with interface dermatitis, allergic contact dermatitis, atopic dermatitis, psoriasis, vitiligo, other diseases in which activated lymphocytes cause damage in the epidermis, skin cancer, and diseases affecting any other tissue known to express the antigens BP180 and/or BP230. In further embodiments, the skin cancer is actinic keratosis, squamous cell carcinoma (SCC), basal cell carcinoma (BCC) or melanoma. In further embodiments, tissues different from skin and mucous membranes that express BP180/BP230 are the retina of the eye (Claudepierre, T et al. J Comp Neurol 2005; 487(2): 190-203) and brain tissue (Seppanen, A et al. Brain Res 2007; 1158:50-6). In some embodiments, the antibody or antigen binding fragment is an scFv.

Monoclonal Antibody

Monoclonal antibodies directed against full length or peptide fragments of a protein or peptide may be prepared using any well-known monoclonal antibody preparation procedures, such as those described for example in Harlow et al. (1988, In: Antibodies, A Laboratory Manual, Cold Spring Harbor, N.Y.) and in Tuszynski et al. (1988, Blood, 72: 109-115).

Quantities of the desired peptide may also be synthesized using chemical synthesis technology. Alternatively, DNA encoding the desired peptide may be cloned and expressed from an appropriate promoter sequence in cells suitable for the generation of large quantities of peptide. Monoclonal antibodies directed against the peptide are generated from mice immunized with the peptide using standard procedures as referenced herein.

Monoclonal antibodies may also be obtained using antibody phage display libraries as previously described (Hammers CM et al. The Journal of investigative dermatology 2014; 134: 1-5; Payne AS et al. J. Clin. Invest. 2005; 115: 888-99).

Nucleic acid encoding the monoclonal antibody obtained using the procedures described herein may be cloned and sequenced using technology which is readily available in the art, and is described, for example, in Wright et al. (1992, Ciritical Rev. Immunol. 12: 125-168), and references cited therein. Further, the antibody of the invention may be“humanized” using the technology described in, for example, Wright et al., and in the references cited therein, and in Gu et al. (1997, Thrombosis and Hematocyst 77:755-759), and other methods of humanizing antibodies well-known in the art or to be developed.

Human Antibody

Provided is a human antibody or antigen binding fragment thereof that binds BP180. In some embodiments, the antibody or antigen binding fragment thereof binds the non-collagenous domain 16A (NC16A) of the BP 180 ectodomain. In further embodiments, the antibody or antibody fragment that binds to non-collagenous domain 16A of BP 180 is listed in Table 3. In yet further embodiments, the antibody or antigen binding fragment thereof comprises 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, and any partial integers therebetween, sequence identity to the amino acid sequence of any one of SEQ ID NOs: 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 61, 62, 63, 64, and 70. Provided is a human antibody or antigen binding fragment thereof that binds BP230. In further embodiments, the antibody or antibody fragment that binds to BP230 is listed in Table 3.

Provided is a human antibody or antigen binding fragment thereof that binds BP180 or BP230, wherein the isolated antibody or antigen binding fragment thereof comprises at least 1, 2, 3, 4, 5 or 6 CDRs from Table 7.

Humanized Antibody

Provided is a humanized antibody or antigen binding fragment thereof that binds BP180. In some embodiments, the antibody or antigen binding fragment thereof binds the non- collagenous domain 16A (NC16A) of the BP180 ectodomain. Also provided is a humanized antibody or antigen binding fragment thereof that binds BP230

Provided is a humanized antibody or antigen binding fragment thereof that binds BP180 or BP230, wherein the isolated antibody or antigen binding fragment thereof comprises at least 1 , 2, 3, 4, 5 or 6 CDRs from Table 7.

Humanized forms of non-human (e g. murine) antibodies are genetically engineered chimeric antibodies or antigen binding fragments thereof having preferably minimal portions derived from non-human antibodies. Humanized antibodies include antibodies in which CDRs of a human antibody (recipient antibody) are replaced by residues from a CDR region of a non human species (donor antibody) such as mouse, rat or rabbit having the desired functionality. In some embodiments, Fv framework residues of the human antibody are replaced by

corresponding non-human residues. Humanized antibodies may also comprise residues that are found neither in the recipient antibody nor in the imported CDR or framework (FR) sequences.

In some embodiments, the humanized antibody may comprise substantially all of at least one, typically two, variable domains domains in which all or substantially ail of the framework regions correspond to those of a relevant human consensus sequence. Humanized antibodies may also include at least a portion of an antibody constant region, such as an Fc region, typically derived from a human antibody (see, for example, Jones et al, 1986. Nature 321 :522-525;

Riechmann et al., 1988. Nature 332:323-329; Presta, 1992, Cum Op. Struct. Biol. 2:593-596). in order to retain high binding affinity, ammo acids in the human acceptor sequence may be replaced by the corresponding amino acids from the donor sequence, for example where: (1) the amino acid is in a CDR; (2) the amino acid is in a human framework region (e.g., the amino acid is immediately adjacent to one of the CDRs). See, U.8. Patent No. 5,530,101 and 5,585,089, incorporated herein by reference, which provide detailed instructions for construction of humanized antibodies.

Although humanized antibodies often incorporate all six CDRs (e.g, as defined by Rabat, but often also including hypervariable loop HI as defined by Chotiua) from a mouse antibody, they can also be made with fewer mouse CDRs and/or less than complete mouse CDR sequence(s) (e.g., a functional fragment of a CDR) (e.g., Pascaiis et al. J Immunol. 169:3076, 2002: Vajdos et al., Journal of Molecular Biology, 320:415-428, 2002; Iwahashi et al, Mol. Immunol. 36: 1079-1091, 1999; Tamura et al., Journal of Immunology!, 164: 1432-1441, 2000).

A humanized antibody has one or more amino acid residues introduced into it from a source which is nonhuman. These nonhuman amino acid residues are often referred to as “import” residues, which are typically taken from an“import” variable domain. Thus, humanized antibodies comprise one or more CDRs from nonhuman immunoglobulin molecules and framework regions from human. Humanization of antibodies is well-known in the art and can essentially be performed following the method of Winter and co-workers (Jones et al, Nature, 321 :522-525 (1986); Riechmann et al., Nature, 332:323-327 (1988); Verhoeyen et al., Science, 239: 1534-1536 (1988)), by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody, i.e., CDR-grafting (EP 239,400; PCT Publication No. WO 91/09967; and U.S. Pat. Nos. 4,816,567; 6,331,415; 5,225,539; 5,530,101 ; 5,585,089; 6,548,640, the contents of which are incorporated herein by reference herein in their entirety). In such humanized chimeric antibodies, substantially less than an intact human variable domain has been substituted by the corresponding sequence from a nonhuman species. In practice, humanized antibodies are typically human antibodies in which some CDR residues and possibly some FR residues are substituted by residues from analogous sites in rodent antibodies. Humanization of antibodies can also be achieved by veneering or resurfacing (EP 592,106; EP 519,596; Padlan, 1991, Molecular Immunology, 28(4/5):489-498; Studnicka et al., Protein Engineering, 7(6):805- 814 (1994); and Roguska et al, PNAS, 91 :969-973 (1994)) or chain shuffling (ET.S. Pat. No. 5,565,332), the contents of which are incorporated herein by reference herein in their entirety.

The choice of human variable domains, both light and heavy, to be used in making the humanized antibodies is to reduce antigenicity. According to the so-called“best-fit” method, the sequence of the variable domain of a rodent antibody is screened against the entire library of known human variable-domain sequences. The human sequence which is closest to that of the rodent is then accepted as the human framework (FR) for the humanized antibody (Sims et al., J. Immunol., 151 :2296 (1993); Chothia et al., J. Mol. Biol., 196:901 (1987), the contents of which are incorporated herein by reference herein in their entirety). Another method uses a particular framework derived from the consensus sequence of all human antibodies of a particular subgroup of light or heavy chains. The same framework may be used for several different humanized antibodies (Carter et al, Proc. Natl. Acad. Sci. USA, 89:4285 (1992); Presta et al, J. Immunol., 151 :2623 (1993), the contents of which are incorporated herein by reference herein in their entirety).

Antibodies can be humanized with retention of high affinity for the target antigen and other favorable biological properties. According to one aspect of the invention, humanized antibodies are prepared by a process of analysis of the parental sequences and various conceptual humanized products using three-dimensional models of the parental and humanized sequences. Three-dimensional immunoglobulin models are commonly available and are familiar to those skilled in the art. Computer programs are available which illustrate and display probable three- dimensional conformational structures of selected candidate immunoglobulin sequences.

Inspection of these displays permits analysis of the likely role of the residues in the functioning of the candidate immunoglobulin sequence, i.e., the analysis of residues that influence the ability of the candidate immunoglobulin to bind the target antigen. In this way, FR residues can be selected and combined from the recipient and import sequences so that the desired antibody characteristic, such as increased affinity for the target antigen, is achieved. In general, the CDR residues are directly and most substantially involved in influencing antigen binding.

A“humanized” antibody retains a similar antigenic specificity as the original antibody. However, using certain methods of humanization, the affinity and/or specificity of binding of the antibody for human CD3 antigen may be increased using methods of“directed evolution,” as described by Wu et al., J. Mol. Biol., 294: 151 (1999), the contents of which are incorporated herein by reference herein in their entirety.

In one embodiment, the antibody is a synthetic antibody, human antibody, a humanized antibody, single chain variable fragment, single domain antibody, an antigen binding fragment thereof, and any combination thereof. Provided is a humanized antibody including a light chain comprising at least one CDR from Table 7 and a human variable region framework; and a heavy chain comprising at least one CDR from Table 7 and a human variable region framework. In some embodiments, the humanized antibody includes said light chain and said heavy chain together with a light chain constant region and a heavy chain constant region.

Hyman or Humanized Single Chain Antibodies

Provided is an isolated human or humanized single chain antibody that binds specifically to BP180 or BP230.

In some embodiments, a human or humanized single chain antibody specific for binding to BP180 or BP230, referred to herein as a“human anti-BP180 single chain antibody,” “humanized anti-BP180 single chain antibody,”“human anti-BP230 single chain antibody” or “humanized anti-BP230 single chain antibody” is fused to an Fc polypeptide. In some embodiments, the Fc polypeptide is an Fc region of an IgG immunoglobulin, such as an IgG immunogl obulin selected from the group consisting of IgG 1 isotype, IgG2 isotype, IgG3 isotype, XgG4 isotype, IgE isotype, IgAl isotype, XgA2 isotype and IgM isotype.

In some embodiments, the human or humanized single chain antibody is fused to the carboxy terminus of the Fc polypeptide. In some embodiments, the human or humanized single chain antibody is fused to the amino terminus of the Fc polypeptide. The fusions are constructed as a single genetic construct and are expressed in cells in culture.

Molecular Biology Techniques

The practice of the present invention employs, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry and immunology, which are well within the purview of the skilled artisan. Such techniques are explained fully in the literature, such as,“Molecular Cloning: A Laboratory Manual”, fourth edition (Sambrook, 2012);“Oligonucleotide Synthesis” (Gait, 1984);“Culture of Animal Cells” (Freshney, 2010);“Methods in Enzymology”“Handbook of Experimental Immunology” (Weir, 1997);“Gene Transfer Vectors for Mammalian Cells” (Miller and Calos, 1987);“Short Protocols in Molecular Biology” (Ausubel, 2002);“Polymerase Chain Reaction: Principles, Applications and Troubleshooting”, (Babar, 2011);“Current Protocols in

Immunology” (Coligan, 2002). These techniques are applicable to the production of the polynucleotides and polypeptides of the invention, and, as such, may be considered in making and practicing the invention. The engineered cytokines of the invention were codon optimized so as to enhance their ability to modulate the immune response in a mammal into which they are introduced. The invention includes sequences that are homologous to the sequences disclosed herein. Sequence homology for nucleotides and amino acids may be determined using FASTA, BLAST and Gapped BLAST (Altschul et al., Nuc. Acids Res., 1997, 25, 3389, which is incorporated herein by reference in its entirety) and PAUP* 4.0b 10 software (D. L. Swofford, Sinauer Associates, Massachusetts). "Percentage of similarity" is calculated using PAUP*

4.0b 10 software (D. L. Swofford, Sinauer Associates, Massachusetts). The average similarity of the consensus sequence is calculated compared to all sequences in the phylogenic tree.

Briefly, the BLAST algorithm, which stands for Basic Local Alignment Search Tool is suitable for determining sequence similarity (Altschul et al, J. Mol. Biol., 1990, 215, 403-410, which is incorporated herein by reference in its entirety). Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information. This algorithm involves first identifying high scoring sequence pair (HSPs) by identifying short words of length W in the query sequence that either match or satisfy some positive-valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as the neighborhood word score threshold (Altschul et al, supra). These initial neighborhood word hits act as seeds for initiating searches to find HSPs containing them. The word hits are extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Extension for the word hits in each direction are halted when: 1) the cumulative alignment score falls off by the quantity X from its maximum achieved value; 2) the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or 3) the end of either sequence is reached. The Blast algorithm parameters W, T and X determine the sensitivity and speed of the alignment. The Blast program uses as defaults a word length (W) of 11, the BLOSUM62 scoring matrix (see Henikoff et al, Proc. Natl. Acad.

Sci. USA, 1992, 89, 10915-10919, which is incorporated herein by reference in its entirety) alignments (B) of 50, expectation (E) of 10, M=5, N=4, and a comparison of both strands. The BLAST algorithm (Karlin et al., Proc. Natl. Acad. Sci. USA, 1993, 90, 5873-5787, which is incorporated herein by reference in its entirety) and Gapped BLAST perform a statistical analysis of the similarity between two sequences. One measure of similarity provided by the BLAST algorithm is the smallest sum probability (P(N)), which provides an indication of the probability by which a match between two nucleotide sequences would occur by chance. For example, a nucleic acid is considered similar to another if the smallest sum probability in comparison of the test nucleic acid to the other nucleic acid is less than about 1 , preferably less than about 0.1, more preferably less than about 0.01, and most preferably less than about 0.001.

Nucleic Acids

Introduction of a nucleic acid encoding any of the engineered antibodies of the invention into a mammal or introduction of a nucleic acid encoding the antibodies or antibody fragments into the cells of a mammal for the purpose of generating antibodies or antibody fragments can be accomplished using technology available in the art.

When taken up by a cell, the genetic construct(s) may remain present in the cell as a functioning extrachromosomal molecule and/or integrate into the cell's chromosomal DNA.

DNA may be introduced into cells where it remains as separate genetic material in the form of a plasmid or plasmids. Alternatively, linear DNA that can integrate into the chromosome may be introduced into the cell. When introducing DNA into the cell, reagents that promote DNA integration into chromosomes may be added. DNA sequences that are useful to promote integration may also be included in the DNA molecule. Alternatively, RNA may be administered to the cell. It is also contemplated to provide the genetic construct as a linear minichromosome including a centromere, telomeres and an origin of replication. Gene constructs may remain part of the genetic material in attenuated live microorganisms or recombinant microbial vectors which live in cells. Gene constructs may be part of genomes of recombinant viral vectors where the genetic material either integrates into the chromosome of the cell or remains

extrachromosomal. Genetic constructs include regulatory elements necessary for gene expression of a nucleic acid molecule. The elements include: a promoter, an initiation codon, a stop codon, and a polyadenylation signal. In addition, enhancers are often required for gene expression of the sequence that encodes the antibody or the immunomodulating protein. It is necessary that these elements be operable linked to the sequence that encodes the desired proteins and that the regulatory elements are operably in the individual to whom they are administered.

Initiation codons and stop codon are generally considered to be part of a nucleotide sequence that encodes the desired protein. However, it is necessary that these elements are functional in the individual to whom the gene construct is administered. The initiation and termination codons must be in frame with the coding sequence. Promoters and polyadenylation signals used must be functional within the cells of the individual.

Examples of promoters useful to practice the present invention, include but are not limited to promoters from Simian Virus 40 (SV40), Mouse Mammary Tumor Virus (MMTV) promoter, Human Immunodeficiency Virus (MV) such as the BIV Long Terminal Repeat (LTR) promoter, Moloney virus, ALV, Cytomegalovirus (CMV) such as the CMV immediate early promoter, Epstein Barr Virus (EBV), Rous Sarcoma Virus (RSV) as well as promoters from human genes such as human Actin, human Myosin, human Hemoglobin, human muscle creatine and human metalothionein.

Examples of polyadenylation signals useful to practice the present invention, include but are not limited to SV40 polyadenylation signals and LTR polyadenylation signals. In particular, the SV40 polyadenylation signal that is in pCEP4 plasmid (Invitrogen, San Diego Calif.), referred to as the SV40 polyadenylation signal, is used.

In addition to the regulatory elements required for DNA expression, other elements may also be included in the DNA molecule. Such additional elements include enhancers. The enhancer may be selected from the group including but not limited to: human Actin, human Myosin, human Hemoglobin, human muscle creatine and viral enhancers such as those from CMV, RSV and EBV.

Genetic constructs can be provided with mammalian origin of replication in order to maintain the construct extrachromosomally and produce multiple copies of the construct in the cell. Plasmids pVAXl, pCEP4 and pREP4 from Invitrogen (San Diego, Calif.) contain the Epstein Barr virus origin of replication and nuclear antigen EBNA-l coding region which produces high copy episomal replication without integration. In order to maximize antibody production, regulatory sequences may be selected which are well suited for gene expression in the cells the construct is administered into. Moreover, codons may be selected which are most efficiently transcribed in the cell. One having ordinary skill in the art can produce DNA constructs that are functional in the cells. In some embodiments for which protein is used, i.e., the engineered antibodies of the invention, for example, one having ordinary skill in the art can, using well known techniques, produce and isolate proteins of the invention using well known techniques. In some embodiments for which protein is used, for example, one having ordinary skill in the art can, using well known techniques, inserts DNA molecules that encode a protein of the invention into a commercially available expression vector for use in well-known expression systems. For example, the commercially available plasmid pSE420 (Invitrogen, San Diego, Calif.) may be used for production of protein in E. cob. The commercially available plasmid pYES2 (Invitrogen, San Diego, Calif.) may, for example, be used for production in S. cerevisiae strains of yeast. The commercially available MAXBAC.TM. complete baculovirus expression system (Invitrogen, San Diego, Calif.) may, for example, be used for production in insect cells. The commercially available plasmid pcDNA I or pcDNA3 (Invitrogen, San Diego, Calif.) may, for example, be used for production in mammalian cells such as Chinese Hamster Ovary cells. For delivery, compounds may also be synthesized as proteins in the patient by introducing DNA expression vectors into that subject (Medi, BM et al. Methods Mol Biol 2008; 423:225-32;

Huang, D et al. Theranostics 2018; 8(9):236l-2376; Todorova, B et al. Sci Rep 2017; 7(l):4l22). One having ordinary skill in the art can use these commercial expression vectors and systems or others to produce protein by routine techniques and readily available starting materials. (See e.g., Sambrook et al, Molecular Cloning, Third Ed. Cold Spring Harbor Press (2001) which is incorporated herein by reference.) Thus, the desired proteins can be prepared in both prokaryotic and eukaryotic systems, resulting in a spectrum of processed forms of the protein.

One having ordinary skill in the art may use other commercially available expression vectors and systems or produce vectors using well known methods and readily available starting materials. Expression systems containing the requisite control sequences, such as promoters and polyadenylation signals, and preferably enhancers are readily available and known in the art for a variety of hosts. See e.g., Sambrook et al, Molecular Cloning Third Ed. Cold Spring Harbor Press (2001). Genetic constructs include the protein coding sequence operably linked to a promoter that is functional in the cell line into which the constructs are transfected. Examples of constitutive promoters include promoters from cytomegalovirus or SV40. Examples of inducible promoters include mouse mammary leukemia virus or metallothionein promoters. Those having ordinary skill in the art can readily produce genetic constructs useful for transfecting with cells with DNA that encodes protein of the invention from readily available starting materials. The expression vector including the DNA that encodes the protein is used to transform the compatible host which is then cultured and maintained under conditions wherein expression of the foreign DNA takes place. The protein produced is recovered from the culture, either by lysing the cells or from the culture medium as appropriate and known to those in the art. One having ordinary skill in the art can, using well known techniques, isolate protein that is produced using such expression systems. The methods of purifying protein from natural sources using antibodies which specifically bind to a specific protein as described above may be equally applied to purifying protein produced by recombinant DNA methodology.

In addition to producing proteins by recombinant techniques, automated peptide synthesizers may also be employed to produce isolated, essentially pure protein. Such techniques are well known to those having ordinary skill in the art and are useful if derivatives which have substitutions not provided for in DNA-encoded protein production.

The polynucleotides encoding the engineered antibodies of the invention may be delivered using any of several well-known technologies including DNA injection, recombinant vectors such as recombinant adenovirus, recombinant adenovirus associated virus and recombinant vaccinia virus.

Routes of administration include, but are not limited to, intramuscular, intranasal, intraperitoneal, intradermal, subcutaneous, intravenous, intra-arterial, intraocular

(intravitreously) and oral as well as topical, transdermal, by inhalation or suppository or to mucosal tissue such as by lavage to vaginal, rectal, urethral, buccal and sublingual tissue.

Preferred routes of administration include intravenous, intramuscular, intraperitoneal, intradermal and subcutaneous injection. Genetic constructs may be administered by means including, but not limited to, electroporation methods and devices, traditional syringes, microneedling devices, needleless injection devices, or microprojectile bombardment.

Antibody-Drug Conjugates

In one aspect, the invention includes a method for treating a disease in a subject in need thereof, the method comprising administering to the subject an effective amount of an antibody- drug conjugate (ADC), wherein the antibody or antigen binding fragment portion of the ADC binds to BP180 or BP230. In some embodiments, the drug used to make the ADC is MMAE (monomethyl auristatin E), ozogamicin, emtansine, amantin, pyrrolobenzodiazepine (PBD) dimer toxin, a chalichaemicin, a cytotoxic maytansinoid, for example DM1.

Provided are antibody-drug conjugates (ADCs) comprising an antibody or antigen binding fragment thereof that bind BP180. Also provided are antibody-drug conjugates (ADCs) comprising an antibody or antigen binding fragment thereof that binds BP230. In some embodiments, the drug is MMAE (monomethyl auristatin E), ozogamicin, emtansine, amanitin, pyrrolobenzodiazepine (PBD) dimer toxin, a chalichaemicin, a cytotoxic maytansinoid, for example DM1.

In some embodiments, the drug is connected to the antibody or antigen binding fragment thereof via a linker. Suitable linkers are known in the art. See for example U/S. Patent No.

8,742,076, which is hereby incorporated by reference in its entirety.

In some embodiments, the antibody portion of the antibody-drug conjugate or the antibody-protein conjugate binds to non-collagenous domain 16A of BP 180 or binds to BP230, and the disease that is treated is bullous pemphigoid, mucous membrane pemphigoid (MMP), linear IgA disease, lichen planus pemphigoides, pemphigoid gestationis, anti-p200 pemphigoid, epidermolysis bullosa acquisita, graft versus host disease (GVHD), toxic epidermolysis bullosa, erythema multiforme, other diseases with interface dermatitis, allergic contact dermatitis, atopic dermatitis, psoriasis, vitiligo, other diseases in which activated lymphocytes cause damage in the epidermis, skin cancer, and diseases affecting any other tissue known to express BP180 and/or BP230 (e.g., ocular diseases or brain diseases). In further embodiments, the skin cancer is actinic keratosis, squamous cell carcinoma (SCC), basal cell carcinoma (BCC) or melanoma. In further embodiments, tissues different from skin and mucous membranes that express BP180/BP230 are the retina of the eye (Claudepierre, T et al. J Comp Neurol 2005; 487(2): 190-203) and brain tissue (Seppanen, A et al. Brain Res 2007; 1158:50-6). Pharmaceutical

The pharmaceutical compositions according to the present invention are formulated according to the mode of administration to be used. In cases where pharmaceutical compositions are injectable pharmaceutical compositions, they are sterile, pyrogen free and particulate free. An isotonic formulation is preferably used. Generally, additives for isotonicity can include sodium chloride, dextrose, mannitol, sorbitol and lactose. In some cases, isotonic solutions such as phosphate buffered saline are preferred. Stabilizers include gelatin and albumin. In some embodiments, a vasoconstriction agent is added to the formulation.

It will be appreciated by a person skilled in the art that the antibody or antigen binding fragment may be administered in admixture with a suitable pharmaceutical excipient diluent or carrier selected with regard to the intended route of administration and standard pharmaceutical practice. For example, see Remington: The Science and Practice of Pharmacy, l9th edition, 1995, Ed. Alfonso Gennaro, Mack Publishing Company, Pennsylvania, USA.

In some embodiments, the antibody or antigen binding fragment may be administered orally, bucally or sublingually in the form of tablets, capsules, ovules, elixirs, solutions or suspensions, which may contain flavoring or coloring agents, for immediate, delayed or controlled-release applications. The antibody or antigen binding fragment may also be administered via intracavernosal injection.

The antibody or antigen binding fragment may also be administered parenterally. In some embodiments, the antibody or antigen binding fragment may be administered

intravenously, intra-articularly, intraperitoneally, intrathecally, intraventricularly, intrasternally, intracranially, intravitreously, intra-muscularly or subcutaneously. In some embodiments, the antibody or antigen binding fragment is administered by infusion techniques.

In some embodiments, the antibody or antigen binding fragment is used in the form of a sterile aqueous solution that may contain other substances, for example, sufficient salts or glucose (or other sugars) to make the solution isotonic with blood. The aqueous solution should be suitably buffered (preferably to a pH of from 3 to 9), if necessary. The preparation of suitable parenteral formulations under sterile conditions is readily accomplished by standard

pharmaceutical techniques well known to a person of skill in the art.

Formulations suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with blood. Suitable formulations for parenteral administration also include aqueous and non-aqueous suspensions which may include suspending agents and thickening agents. The formulations may be presented in unit-dose or multi-dose containers.

For parenteral administration to human patients, the daily dosage level of the antibody or antigen binding fragment that binds BP180 or BP230 will usually be from 10 to 200 g per adult (i.e., from about 0.1 to 2 g/kg), administered in single or multiple or divided doses.

In some embodiments, the dosage level may be from about 1 mg/kg to about 10 mg/kg, depending on the site and exact route of application, e. g., intraocular injection or topical administration.

In some embodiments, the antibody or antigen binding fragment is administered intranasally or by inhalation. The antibody or antigen binding fragment may be delivered in the form of a dry powder inhaler or an aerosol spray from a pressurized container, pump, spray or nebulizer with the use of a propellant.

In some embodiments, the antibody or antigen binding fragment is administered by DNA injection and electroporation of the DNA encoded antibody into muscle or skin.

Methods of Treatment

Provided is a method of treating a disease in a subject in need thereof, comprising administering to the subject an effective amount of any one of the antibody or antigen binding fragments described herein. In some embodiments, the subject is human. In some embodiments, the antibody or antigen binding fragment is provided in a pharmaceutical composition.

In some embodiments, the pharmaceutical composition may be delivered orally, parenterally, for example as a parenteral injection, intravenously, for example as an intravenous infusion, or by inhalation.

In some embodiments, a sample is a tissue or a bodily fluid sample. In some

embodiments, the sample is a tumor sample, a blood sample, a blood plasma sample, a peritoneal fluid sample, an exudate or an effusion.

In some embodiments, a second agent is administered to the subject. In further embodiments, the second agent is at least one of carboplatin, cisplatin, paclitaxel, docetaxel, gemcitabine, bevacizumab, olaparib, rucaparib, niraparib, cyclophosphamide, FU, abiraterone, flutamide, bicalutamide, leuprolide, goserelin, buserelin, triptorelin, degarelix, Enzalutamide, Apalutamide, Sipuleucel-T, Cabazitaxel, Radium-223, trastuzumab, pertuzumab, lapatinib, tamoxifen, oxaliplatin, capecitabine, leucovorin, Irinotecan, Cetuximab, panitumumab, aflibercept, Regorafenib, Trifluridine-tipiracil, immune checkpoint inhibitors (nivolumab, pembrolizumab), cabozantinib, sunitinib, pazopanib, axitinib, interleukin-2, interferon alpha, mitomycin C, epirubicin, BCG, bleomycin, etoposide, sorafenib, regorafenib, rituximab, dupilumab, efgartigimod, coversin, lenvatinib, pemetrexed and/or vinorelbine. In some embodiments, the second agent is an alkylating agent, antimetabolite, antibiotic, a plant-derived agent, platinum complex, campthotecin derivative, tyrosine kinase inhibitor, monoclonal antibody, interferon, biological response modifier, hormonal anti-tumor agent, anti-tumor viral agent, angiogenesis inhibitor, differentiating agent, PI3K/mTOR/AKT inhibitor, cell cycle inhibitor, apoptosis inhibitor, hsp 90 inhibitor, tubulin inhibitor, DNA repair inhibitor, anti- angiogenic agent, receptor tyrosine kinase inhibitor, topoisomerase inhibitor, taxane, agent targeting Her-2, hormone antagonist, agent targeting a growth factor receptor, or a

pharmaceutically acceptable salt thereof. In some embodiments, the anti-tumor agent is citabine, capecitabine, valopicitabine or gemcitabine. In some embodiments, the anti-tumor agent is Avastin, Sutent, Nexavar, Recentin, ABT-869, Axitinib, Irinotecan, topotecan, paclitaxel, docetaxel, lapatinib, Herceptin, tamoxifen, progesterone, a steroidal aromatase inhibitor, a non steroidal aromatase inhibitor, Fulvestrant, an inhibitor of epidermal growth factor receptor (EGFR), Cetuximab, Panitumimab, an inhibitor of insulin-like growth factor 1 receptor (IGF1R), and/or CP-751871.

In some embodiments, the disease is cancer. In further embodiments, the cancer is skin cancer. In some embodiments, the skin cancer is actinic keratosis, squamous cell carcinoma (SCC), basal cell carcinoma (BCC) or melanoma.

In some embodiments, the antibody or antigen binding fragment binds to non- collagenous domain 16A of BP180, and the disease is bullous pemphigoid, mucous membrane pemphigoid (MMP), linear IgA disease, lichen planus pemphigoides or pemphigoid gestationis, or any other disease affecting tissues that express BP180/BP230.

Antibody- Cell-Mediated Cytotoxicity (ADCC) Antibody-dependent cell-mediated cytotoxicity (ADCC) is a cell-mediated reaction in which nonspecific cytotoxic cells that express Fc receptors (FcRs), such as natural killer (NK) cells, neutrophils and macrophages, recognize bound antibody on a target cell and subsequently cause lysis of the target cell. The primary cells for mediating ADCC, NK cells, express FcgRIII only, whereas monocytes express FcgRI, FcgRII and FcgRIII. FcR expression on hematopoietic cells is summarized in Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991). To assess ADCC activity of a molecule of interest, an in vitro ADCC assay, such as that described in U.S. Patent Nos. 5,500,362 or 5,821,337 may be performed. Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC) and Natural Killer (NK) cells.

Alternatively, or additionally, ADCC activity of the molecule of interest may be assessed in vivo, e.g., in an animal model.

In some embodiments, administering to the subject an effective amount of any one of the antibody or antigen binding fragments described herein results in ADCC.

EXPERIMENTAL EXAMPLES

The invention is further described in detail by reference to the following experimental examples. These examples are provided for purposes of illustration only, and are not intended to be limiting unless otherwise specified. Thus, the invention should in no way be construed as being limited to the following examples, but rather, should be construed to encompass any and all variations which become evident as a result of the teaching provided herein.

Without further description, it is believed that one of ordinary skill in the art can, using the preceding description and the following illustrative examples, make and utilize the compounds of the present invention and practice the claimed methods. The following working examples therefore, specifically point out the preferred embodiments of the present invention, and are not to be construed as limiting in any way the remainder of the disclosure.

Materials and Methods

Patients and serum samples

Following the principles of the Declaration of Helsinki, sera for immunofluorescence testing were obtained from active BP patients with typical clinical presentation and positive test results in BP180-NC16A and/or BP230-C/N IgG ELISA testing (MBL, Nagoya, Japan) after obtaining written informed consent and Institutional Review Board approval. Sera were tested on MBL kits because here both the BP230-C- and the BP230-N-terminus are available as substrates, and negative BP230 titers on this ELISA system indicates absence of both BP230-C and BP230- N reactivity; EUROIMMUN only uses a BP230-C fragment as substrate (Hamada T et al.

Experimental dermatology 2001; 10: 256-63; Blocker IM et al. Epitope mapping of BP230 leading to a novel enzyme-linked immunosorbent assay for autoantibodies in bullous

pemphigoid. The British journal of dermatology 2012; 166: 964-70). Lor initial full serum immunofluorescence studies, 10 patients with concurrent BP180-NC16A and BP230-C/N IgG reactivity, 7 patients with BP230-C/N (but no BP180-NC16A) IgG reactivity, and 12 patients with BP180-NC16A (but no BP230-C/N) IgG reactivity (all by MBL ELISA testing according to the manufacturers’ instructions) were included. Lor subsequent affinity purification of antigen- specific antibodies and related immunofluorescence tests, an additional 2 sera with high reactivities against both BP AGs were included.

Indirect immunofluorescence (IIF) of patient sera

Normal human skin (NHS) was acquired from dermatologic surgery procedures using Institutional Review Board-reviewed protocols and cryosectioned onto glass slides (5 pm thickness). ME sections were obtained from SCIMEDX (Dover, NJ, USA). Slides were blocked with TBS-Ca ²⁺ /l% BSA at RT for 30 min. Slides were washed with TBS-Ca ²⁺ three times and incubated with serum samples diluted 1 : 100 (and, additionally, 1 : 10 in cases of negative immunofluorescence results at 1 : 100) in TBS-Ca ²⁺ /l% BSA at RT for 60 minutes. Binding of bound antibodies was detected with anti-human IgG F(ab’)2 FITC conjugate (dilution, 1 : 100; Bio-Rad). Finally, slides were washed as above and mounted with DAPI Fluoromo20unt-G mounting medium (SouthernBiotech).

Affinity purification of BP AG-specific antibodies from human sera and IIF

Recombinantly produced, His-tagged BP180-NC16A and BP230-C fragments were obtained from EUROIMMUN and coupled to magnetic beads (Dynabeads™ His-Tag Isolation & Pulldown; ThermoFisher Scientific), following the instructions of the manufacturer. Serum was diluted 1 : 10 diluted in DPBS/0.05% Tween 20, pH 7.1, and incubated with antigen-loaded beads for 90 minutes. After extensive washing with DPBS/0.05% Tween 20, pH 7.1, antigen- specific antibodies were eluted with 76 mM citric acid, pH 2.2, and immediately neutralized with 2M TRIS buffer, pH 11.1. Original BP sera, eluates of antigen-specific affinity purifications, and the sample flow-throughs were analyzed for complete affinity purification by ELISA testing (EUROIMMUN), according to manufacturers’ directions and adjusted to the respective original serum concentrations used. IIF for bound IgG was performed on ME substrates (SCIMEDX; EUROIMMUN), adjusting samples to the respective original input serum concentrations and using above IIF protocol.

Isolation of monoclonal and scFv antibodies from a BP patient

Using previously described methods (Hammers CM el al. The Journal of investigative dermatology 2014; 134: 1-5; Payne AS et al. J. Clin. Invest. 2005; 115: 888-99), antibody phage display (APD) libraries were constructed from peripheral mononuclear cell-RNA, obtained from a patient with active BP (confirmed by clinical features, DIF, IIF, ELISA). In brief, the PCR- amplified variable heavy and light chains were assembled via overlap-PCR, digested with Sfi-I (Roche), and ligated into the pComb3X vector (Scripps Institute, La Jolla). After electroporation into XLl-Blue (Agilent), phage libraries were panned on BP180/230-ELISA wells (MBL, Nagoya, Japan). Bound polyclonal phages were eluted with 76 mM citric acid, amplified, and re panned for up to four rounds. All isolated monoclonal antibodies (mAbs; in form of single chain variable fragments, scFv) were sequenced, individually re-tested on BP180/230 ELISA substrates (MBL, Nagoya, Japan, and EUROIMMUN, Luebeck, Germany) for binding, using an anti -Ml 3 ELISA (for monoclonal phage) and an anti-HA-ELISA (for HA-tagged scFvs, after expression in TOP10F’ E. coli, Invitrogen). Because both BP230 clones used herein reacted on both the MBL BP230-C/N and the EUROIMMUN BP230-CF-ELISA systems, we concluded that both clones react against the BP230-C domain (clones termed BP230-C-mAbl [4-8E, scFv# 28; Table 3] and BP230-C-mAb2 [4-8G, scFv# 14; Table 3]). Monoclonals against BP180- NC16A used herein were termed BPl80-NCl6A-mAbl (3-4E scFv# 17; Table 3) and BP180- NCl6A-mAb2 (3-14G, scFv# 1; Table 3).

Monoclonal IIF

NHS cryosections of 6 pm thickness were blocked with TBS-Ca (Bio-Rad) plus 1% BSA (Sigma-Aldrich) at RT for 30 min. Slides were washed with TBS-Ca ²⁺ three times and incubated with samples diluted in TBS-Ca ²⁺ plus 1% BSA at RT for 60 minutes. Binding of scFvs was detected through staining with rat anti -HA mAb (3F10; dilution, 1 : 100; Roche), followed by an Alexa Fluor 488-conjugated anti-rat IgG (dilution, 1 :200; Invitrogen). Analogously, IIF was performed on ME substrates (SCIMEDX; EUROIMMUN), following above protocol.

Double IIF

For double IIF, the above detailed protocols for IIF staining of affinity-purified polyclonal IgG (dilution 1 :40 each) and monoclonal scFv (dilution 1 : 1,000) were serially executed on the same tissue substrates.

Displacement Experiments

Displacement of serum IgG by scFv was studied using the following protocol. Patient serum was incubated with NCl6A-wells (5 wells) plus one well without serum (here: blocking buffer) for 30 minutes at 37 degrees Celsius. The patient serum was titered before, for an OD of about 0.5/linear range. After one wash, scFvs #1, 29, 16 and 28 in different dilutions (stock at about 1 ug/uL): 1 : 10, 1 : 100, 1 : 1,000, 1 : 10,000. 2 wells were without scFv (positive control with serum IgG only; negative control). The wells were incubated for 2 hours at 37 degrees Celsius. After two washes, the previous step was repeated with fresh scFv dilutions and the wells were incubated 30 minutes at 37 degrees Celsius (equals re-infusion/re-injection in patients). After two washes, the wells were incubated with anti-IgG-HRP (from EUROIMMUN kit) for 30 minutes at room temperature. After three washes, the wells were developed for 10 minutes and were read out at 450 nm wavelength. Optical density (OD) values are given in Tables 4-6.

Sequence analyses and alignments

All sequence analyses are based on publicly accessible sequence data at NIH NCBI BLAST®, available online at blast.ncbi.nlm.nih.gov (Protein BLAST®; last accessed on 04/19/18, 11 :58 PM). Alignments are based on NIH NCBI BLAST®, EMBL-EBI Clustal Omega (at www.ebi.ac.uk/Tools/msa/clustalo/) and SnapGene® Version 4.1.4 for Mac (GSL Biotech LLC, Chicago, IL, USA). scFv Sequences

3-14G SEQ ID NO: 1

GAGCTCACACTCACGCAGTCTCCAGGCACCCTGGCTTTGTCTCCAGGGGAAAGAGCC

ACCCTCTCCTGCAGGGCCAGTCAGAGTGTTAGCAGCAACTACTTAGCCTGGTACCAG

CAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGGTGCATCCAGCAGGGCC CCT GGC AT CCC AGAC AGGTT C AGTGGC AGT GGGT CT GGGAC AGATTT C ACT CT C ACC

ATCACCAGCCTGCAGGCTGAAGATGTGGCAGTTTATTACTGTCAGCAATATTTTGAT

AGTCCGCTCACTTTCGGCGGAGGGACCAAGGTGGAAATCAAAGGTGGTTCCTCT

AGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGCAGGTGCAGCTGGTG

CAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTCTCCTGCAAGGC

TTCTGGATACACCTTCAGCAGTTATGACATCAATTGGGTGCGACAGGCCACTGGA

CAACGGCCTGAGTGGATGGGGTGGATGAACCCTAAAAGTGGTGACACAGGCTACGC

ACAGACCTTCCAGGGCAGAGTCACCATGACCAGGAGCACCTCCATAAGTACAGCCT

ATATGGAACTGAGCAGCCTGAGATCTGAAGACACGGCCATGTATTATTGTACGAGA

GGCAACCGACCGATTTTTGATTTATTTCCTTTTGATCTCTGGGGCCAAGGGACACTG

ATCACCGTCTCTTCA

3-17G SEQ ID NO: 2

GAGCTCGTGCTGACTCAGCCACCTTCAGCGTCTGGGACCCCCGGACAGAGGGTCAC

CATCTCTTGTTCTGGAAGCAGCTCCAACATCGGACTTAATTTTGTATACTGGTATAT C

CAGTTCCCGGGAACGGCCCCCAAACTCCTCATCTATAGGAATAATCAGCGGCCC

TCAGGGGTCCCTGACCGAATCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCC

ATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTACATCATGGGATGCC

AGCCTGAGTGCTTGGGTGTTCGGCGGAGGCACCAAGCTGACCGTCCTAGGCGGT

GGTTCCTCTAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGCAGGTG

CAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCGGAGACCCTGTCCCTCAC

CTGCACTGTCTCTGGTGGCTCCGTCAGCAGTGGTAGTTACTACTGGAGCTGGATC

CGGC AGCCCCC AGGGAAGGGACT GGAGT GGATT GGGAGT AT GTTTT AT AGTGGGAG

AACCTACTACAACCCGTCCCTCAACAGTCGAGTCACCATATCCGGAGACACGTCCAA

GAACCAGGTCTCCCTGAAGCTGAGCTCTGCGACCGCCACAGACACGGCTGTATAT

TACTGTGCGAGACGCGGGAGCCGACTAAATACTGTTGGAGTTCCACCTGCTATTGAA

CTCTGGGGCCAGGGAACCCTGGTCACCGTCTCCCCA

3-30G SEQ ID NO: 3

GAGCTCGTGCTGACTCAATCGCCCTCAGCGTCTGCGGCCCCCGGGCAGAGGATCACC

ATCTCTTGTTCTGGAAGCAGCTCCAACATCGGAATTAATTATGTATACTGGTACCAG CAGGTCCCAGGAACGGCCCCCAAACTCCTCATCTATAGGAATAATCAGCGGCCCTC

AGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCCGGCACCTCAGCCTCCCTGGCCAT

CAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGCAGCATGGGATGACA

GCCTGGATGGTCATTGGGTGTTCGGCGGAGGGACCAAGGTGACCGTCCTAGGTGGT

GGTTCCTCTAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGCAGGTG

CAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCGGAGACCCTGTCCCTCAC

CTGCACTGTCTCTGGTGGCTCCATCAGCAGTGGTGGTTACTACTGGAGCTGGATCCG

CCAGCCCCCAGGGAAGGGGCTGGAGTGGATTGGGAGTATCTTTTATAGTGGGAGAA

CCTACTACAACCCGTCCCTCAACAGTCGAGTCACCATATCCGGAGACACGTCCAAGA

ACCAGGTCTCCCTGAAGCTGAGCTCTGTGACCGCCACAGACACGGCTGTATATTACT

GTGCGAGACGAGGAACCCGTCTCAGTGAAACAGTGGCCCCGGCCTATGAGTACTGG

GGCCAGGGAACCCTGGTCACCGTCTCCTCA

3-4G SEQ ID NO: 4

GAGCTCGTGCTGACTCAATCGCCCTCAGCGTCTGGGACCCCCGGGCAGAGGGTCAC

CAT CTCTT GTTCTGGAAGC AGCT CCA AC AT CGGAAGTT ATTTT GT AT ATT GGT AT C AG

CAACTCCCAGGAACGGCCCCCAAACTCCTCATCTATAGGAATAGTCAGCGGCCCTCA

GGGGTCTCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATC

AGTGGGCTCCAGTCTGAGGATGAGGCTGATTATTACTGTGCATCATGGGATGACAGC

CTGAGTGCTTGGGTTTTCGGCGGAGGGACCCAGCTGACCGTCCTCGGTGGTGGTTCC

TCTAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGCAGGTGCAGCTG

CAGGAGTCGGGCCCAGGACCGGTGAAGCCTTCGGAGACCCTGTCCCTCACCTGCAC

TGTCTCTGGTGGCTCCATCAGCAGTAATACTTACTACTGGGAGTGGATCCGCCAGCC

CCCAGGGAAGGGGCTGGAGTGGATTGGGAGTATGTTTTATAGTGGGAGAACCTACT

ACAACCCGTCCCTCAACAGTCGAGTCACCATATCCGGAGACACGTCCAAGAACCAG

GTCTCCCTGAAACTAAGCTCTGTGACCGCCGCGGACACGGCTGTGTATTACTGTGCG

AGACGATCTTTACGATATTTTGACTGGTCCTATGACTACTGGGGCCAGGGAACCCTG

GTCACCGTCTCCTCA

3-29G SEQ ID NO: 5 GAGCTCGGGCAGACTCAGCAGCTCTCAGCGTCTGGGACCCCCGGGCAGAGGGTCAC

CATCTCTTGTTCTGGAAGCAGCTCCAACATCGGAAGTCATTATGTATACTGGTACCA

GCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATAGGAATAATCAGCGGCCC

CCAGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGCCACCTCAGCCTCCCTGGCC

ATCAGTGGGCTCCAGTCTGAGGATGAGGCTGATTATTACTGTGCAGCATGGGATGAC

AGCCTGGATGGTCATTGGGTGTTCGGCGGAGGCACCCAGCTGACCGTCCTCGACGGT

GGTTCCTCTAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGCAGGTG

CAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCCTCGGAGACCCTGTCCCTCAC

CTGCACTGTCTCTGGTGGCTCCATCAGCAGTGATACTTACTACTGGGACTGGATCCG

CC AGCCCCC AGGGAGGGGGCT GGAGT GGATTGGC AGT AT C AATT ATAGTGGGAAC A

CCTACTACAACCCGTCCCTCAAGAGTCGAGTCACCATATCCGTAGACACGTCCAAGA

ACCAGTTCTCCCTGAAGCTGAGCTCTGTGACCGCCGCAGACACGGCTGTGTATTACT

GTGCGAGACTGGGTAGCAGTGGCTGGTACTTGCCTTTTGACTACTGGGGCCAGGGAA

CCCTGGTCACCGTCTCCTCA

3-3G SEQ ID NO: 6

GAGCTCGAGCTGACTCAGCCACCCTCAGTGTCTGGGACCCCCGGGCAGAGGGTCAC

CATCTCTTGTTCTGGCAGCAGCTCCAGCATCGGAAGTAATACTGTAGACTGGTACCA

GCAGGTCCCAGGAACGGCCCCCAAACTCCTCATCTATAGTAATAATCAGCGGCCCTC

AGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCAT

CAGTGGGCTCCAGTCTGAGGATGAGGCTGATTATTACTGTGCAGCATGGGATGACA

GCCTGAATGGTTATGTGCTATTCGGCGGAGGGACCAAGCTGACCGTCCTAGGCGGTG

GTTCCTCTAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGCAGGTGC

AGCT GGT GC AGTCTGGGGCT GAGGT GA AGAAGCCTGGGGCCT C AGT GAAGGT CTCC

TGCAAGGCCTCTGGATACACCTTCACCGACCACTATATACACTGGGTGCGACAGGCC

CCT GGAC AAGGGCTT GAGT GGAT GGGAT GGAT C AACCCT AAC AGT GGT GCC AC AAA

TC AT GC AC AGAGGTTTC AGGGC AGGGT CT CC ATGACCT GGGAC ACGT CC AGC AGC A

CAGTCAACATGGAGCTGAGCAGGCTGAGATCTGACGACACGGCCGTGTATTACTGT

GCGAGAGGACCCTACATTAACTACGACTACTGGGGCCAGGGAACCCTGGTCACCGT

CTCCTCA 3-9G SEQ ID NO: 7

GAGCTCGTGCTGACTCAGCCACCTTCAGCGTCTGGGACCCCCGGGCAGAGGGTCAC

CATCTCTTGTTCTGGAAGCAGCTCCAACATCGGAAGTAATTATGTATACTGGTACCA

GCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATAGGAATAATCAGCGGCCC

TCAGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCC

ATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGAGCGTGGGATGG

CAATCTGCATGCTTGGGTCTTCGGCGGAGGGACCAAGCTGACCGTCCTAGGTGGT

GGTTCCTCTAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGCAGGTG

CAGCTGGTGCAGTCTGGGACTGAAGTGAAGAAGGCTGGGGCCTCAGTGAAGGTTTC

CTGCAAGGCATCTGGAAACACCTTCACCAGCTACTATATGCACTGGGTGCGACAG

GCCCCTGGACAAGGGCTTGAGTGGATGGGAGTAATCGACCCTAGTGGTGGTAGCAC

AACCTACGCACAGAAGTTTCAGGGCGGAGTCACCATGACCAGGGACACGTCCACGA

GCGCAGTCTACATGGAGTTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTAC

TGTGCGAGAGGGAGGCTTCGGGGAGTTATTATGAGTCCATTTGACTACTGGGGCCAG

GGAACCCTGGTCACCGTCTCCTCA

3-12G SEQ ID NO: 8

GAGCTCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTC

ACCATCACTTGCCGGGCCAGTCAGGGTATTGCTAACTGGTTGGCCTGGTATCAGCAG

AAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGG

GTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGAATTCACTCTCTCCATCAGC

AGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACACTATCACAGTTATCCG T

ACACTTTTGGCCAGGGGACCAAGCTGGAGATCAAAGGTGGTTCCTCTAGATCTTCCT

CCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGCAGGTGCAGCTGGTGCAGTCTGGG

GCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTTTCCTGCAAGGCATCTGGATA

CACCTTCAGCAACTACTATATGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTTG

AGTGGAT GGGAAT AAT C AACCCT AGT GGT GGT GGC AC AAGAT ACGC AGAGACGTT C

CAGGGCAGAATCACCATGACCAGGGACACGTCCACGAACACAGTCTACATGGAGTT

GAGC AGTTT GAGAT CT GAGGAC ACGGCCGT AT ATT ATT GT GCGCGAGAT GT GGAGG

CCTATGGTTCTTTTATGGCCTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA 3-27G SEQ ID NO: 9

GAGCTCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTATAGGAGACAGAGTC

ACCATAGCCTGCCGGGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAG

AAGCCAGGGAAAGCCCCTAAGCTCCTGATCTATGATGCATCCAGTTTGCAAAGTGG

GGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAG

CAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACAGTGTACC

GCTCACTTTCGGCGGAGGGACCAAGGTGGAAATCAAAGGTGGTTCCTCTAGATCTTC

CTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGCAGGTGCAGCTGGTGCAGTCTGG

AGCTGAGGTGAGGAAGCCTGGGGCCTCAGTGAAGGTCTCCTGCGAGGCTTCTGGAT

ACACCCTCACCAACTACTATATGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTT

GAGT GGAT GGGAGT A AT C A AC CC T AGT GGT GGT AGT AC A A AC T ACGC AC AGA AGTT

CC AGGGC AGAGTC ACC AT GACT AGGGAC ACGTCC ACGAGC AC AGT CT AT ATGGAGC

TGAGCAGCCTGAGACCTGAGGACACGGCCATATATTACTGTGCGAGAGATATGAGA

CCGAATCACTATGCTTCGGGGAGTTTTAACTACTGGGGCCAGGGAACCCTGGTTACC

GTCTCCTCA

3-5G SEQ ID NO: 10

GAGCTCACACTCACGCAGTCTCCAGGCACCCTGTCTTTGTCTCCAGGGGAAAGAGCC

ACCCTCTCCTGCAGGGCCAGTCAGAGTGTTAACAACAAGTATTTAGCCTGGTACCAG

CAGAAGCCTGGCCAGGCTCCCACGCTCCTCATCTATGGTGGATCCATGAGGGCC

ACCGGCATCCCGGACAGGTTCAGTGGCAGTGGGTCTGAGACAGACTTCACTCTCACC

ATCACCAGACTGGAGCCTGAAGATTTTGCTGTGTATTACTGTCAGCAGTATGGTAGG

TCACCTATCACCTTCGGCCAAGGGACACGACTGGAGATTAAAGGTGGTTCCTCTAGA

TCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGCAGGTGCAGCTGGTGCAG

TCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCTTCT

GGAGGC ACCTT C AGC AACT CT GCT AT C AGTT GGGTGCGAC AGGCCCCTGGAC AAGG

GCTTGAGTGGATGGGAGCAATCATCCCTATCTTTGGTACACCAAACTACGCACAGAG

ATTCCAGGGCAGAGTCACGATTACCGCGGACGAATCCACGAGAACAGCCTACATGG

AGCTGGGCAGCCTGAGATCTGAGGACACGGCCGTTTATTACTGCGCGAGAGATCAA

CCGTCGTGGGGCAGTGGCTGGTCCCCAGACCACTACTACGGTCTGGACGTCTGGGGC

C AAGGGACC ACCGT C ACCGTCTCCT C A 3-34G SEQ ID NO: 11

GAGCTCGTGCTGACTCAATCGCCCTCAGCGTCTGGGACCCCCGGGCAGAGGGTCAC

CATCTCTTGTTCTGGGAGCAGCTCCAACATCGGAGGTAACTATGTATACTGGTTCCA

GCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATAGGAATAATCAGCGGCCC

TCAGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCC

ATCAGTGGGCTCCAGTCTGAAGATGAAGGTGATTATTACTGTGCAGCATGGGATGCC

AGCCTGAAACATCGACTGTTCGGCGGAGGCACCAAGGTGACCGTCCTAGGCGGTGG

TTCCTCTAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGGAGGTGCA

GCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTG

TTCAGCCTCTGGATTCACCTTTAGTAACTATTGGATGAGCTGGGTCCGCCAGGCTCC

AGGGAAGGGGCTGGAGTGGGTCTCATCCATTAGTAGTAGTAGTAATTACATATACTA

CGCGGACT C AAT GAAGGGCCGATT C ACC AT CTCC AGAGAC AACGCC A AGAACT C AC

TGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCG

AGAGTCTATAGCAGGTGGCTTAGGTACTTTGAGTACTGGGGCCAGGGAACCCTGGTC

ACCGTCTCCTCA

4-4G.230 SEQ ID NO: 12

GAGCTCGTGTTGACACAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTC

ACC ATCCCTT GCCGGGC A AGT C AC AAC ATT AAAAACT ATTT AA ATT GGT AT C AGC AG

AAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATACACTTTGCAAGGT

GGAGT CCC AT C AAGGTT C AGT GGC AGT GGATCTGGGAC AGATTT C AAT CT C ACC AT C

AGCAGCCTGCAGCCTGAAGATTTTGCAACTTACTACTGCCAACAGAGTTACAGTACC

CCTCCGGCTTTCGGCGGCGGGACCAAAGTGGATATCAAAGGTGGTTCCTCTAGA

TCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGGAGGTGCAGCTGGTGGAG

TCTGGGGGAAACTTGGTACAGCCGGGGGGGTCCCTGAGACTCTCCTGTTCAGCCTCT

GGATTC ACCTTT GAGGATT AT GCC ATGC ACT GGGT CCGGC AAGCT CC AGGGAAG

GGCCTGGAGTGGGTCGCGGGTATATTTTGGAATAGCGACAGCATAGGCTATGGGGA

CTCTGTGAAGGGCCGATTCACCATCTCCAGAGACAACGCCAAGAACTCCCTGTATCT

GCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCGAGAGTA GGTTACGATTTTTGGAGTGGTTATTCAACCCCCTTTGACTACTGGGGCCAGGGAACC

CTGGTCACCGTCTCCTCA

4-5G.230 SEQ ID NO: 13

GAGCTCGTGCTGACTCAGCCACCTTCAGTGTCTGGGCCCCCCGGGCAGAGGGCCACC

ATCTCTTGTTCTGGAAGCAGCTCCAACTTCGGAAGTTATAATGTAAACTGGTACCAG

CAGCTCCCGGGAACGGCCCCCAAACTCCTCATCCATAATAATAATCAGCGGCCC

TCAGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCC

ATC AGT GGGCT CC AGT CT GAGGAT GAGGCT GAGT ATTACTGT GC AGT GT GGT ATGAC

AGTCTTAATGGTCCGGTATTCGGCGGAGGCACCCAGCTGACCGTCCTCGGCGGTGGT

TCCTCTAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGCAGGTGCAG

CTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGC

AAGGCTTCTGGAGGCACCTTCAACACCTATCCTTTCACCTGGGTACGGCAGGCCCCT

GGACAACGGCTTGAGTGGGTGGGAGAAATCACCCCTCTCTTTGGAACAGCAGACTA

CGCACAGAGGTTCCAGGGCAGAGTCACGATAAGCGCGGACGAACCCACGACCACTG

TTTATATGGAGCTGAACAACCTGAGATCTGAAGACACGGCCGTATATTTCTGTATGA

CCCCTTTTACCCACTCCTACTACTACGGAATGGACGCCTGGGGCCAAGGGACCACGG

TCACCGTCTCCTCA

4-8G.230 SEQ ID NO: 14

GAGCTCACACTCACGCAGTCTCCAGGCACCCTGTCTTTGTCGCCAGGGGAAAGAGCC

ACCCTCTCCTGCAGGGCCAGTCAGAATATTAACAGCAGGTTAGCCTGGTACCAGCA

GAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGGTGCATCCACCAGGGCCACTGG

TATCCCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACTCTCACCATCAG

CAGACTGGAGCCTGAAGATTTTGCAGTGTATTACTGTCAGCACTATGGTAGCTCACC

TCAGCTCAGGACTTTTGGCCAGGGGACCAAGCTGGAGATCAAAGGTGGTTCCTCTAG

ATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGCAGGTGCAGCTGGTGCA

GTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCTTC

TGGAGGCACCTTCAACACCTATCCTTTCACCTGGGTACGGCAGGCCCCTGGACAACG

GCTTGAGTGGGTGGGAGAAATCACCCCTCTCTTTGGAACAGCAGACTACGCACAGA

GGTTCCAGGGCAGAGTCACGATAAGCGCGGACGAACCCACGACCACTGTTTATATG GAGCTGAACAACCTGAGATCTGAAGACACGGCCGTATATTTCTGTACGACCCCTTTT

ACCCACTCCTACTACTACGGAATGGACGCCTGGGGCCAAGGGACCACGGTCACCGT

CTCCTCA

3-47E SEQ ID NO: 15

GAGCTCGAGCTGACTCAGCCACCCTCAGTGTCCGGGTCTCCTGGACAGTCAGTCACC

ATCTCCTGCACTGGAACCAGCAGTGACGTTGGTGGTTATAAGTATGTCTCCTGGTAC

CAACAACACCCAGGCGAATCCCCCAAACTCTTGATTTATGAGGCCAGGAAGCGGCC

CTCAGGGGTCCCTGATCGCTTCTCTGGCTCCAAGTCGGGCAAGACGGCCTCCCTGAC

TGTCTCTGACCTCCAGGCTGACGATGAGGCTTATTATTATTGCAGCTCATATGGAGG

CAGCGACAATGTACTATTCGGCGGAGGGACCAAGGTGACCGTCCTAGGCGGTGGTT

CCTCTAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGGAGGTGCAGC

TGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGACGGTCTCCTGC

ACGGCTTCTGGATACACCTTCAGCAGTTATGATATCAACTGGGTGCGACAGGCCACT

GGACAAGGGCCTGAGTGGATGGGATGGATGAACCCTAAGAGTGGTGAAACCGGCTA

TGCACAGAAGTTCCAGGGCAGAGTCACCATGACCAGGAACACCTCCATAAGAACAG

CCTACTTGGAGTTGAGCAGCCTGAGATCTGACGACACGGCCATATATTATTGTGCGA

GAGGCAACCGACCAATTTTTAATTTGTTCCCTTTTGATCTGTGGGGCCAGGGGACAA

TGGTCACCGTCTCTTCA

3-3E SEQ ID NO: 16

GAGCT CGAGCT GACTC AGCC ACCCT C AGT GT CT GGGGCCCC AGGGC AGAGGGT C AC

CATCTCCTGCACTGGGAGCAGCTCCAACATCGGGGCAGGTTATCATGTACACTGGTA

TCAGCAGTTTCCGGGAACAGCCCCCAAACTCCTCATTTATGATAACAACAATCGGCC

CTCAGGGGTCCCTGATCGATTCTCTGGCTCCAAGTCTGGCGCTTCAGCCTCCCTGGC

CAT C ACTGGGCT CC AGGCTGAT GAT GAGGCTGATT ATT ACT GCC AGT CCT AT GAC AG

CAGCCTGAGTGGGAATTGGGTGTTCGGCGGAGGCACCGAGCTGACCGTCCTCGGCG

GTGGTTCCTCTAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGCAGG

TGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAAGATC

TCCTGTAAGGGTTCTGGATACAACTTTAGTAACTACTGGATCGGCTGGGTGCGCCAG

ATGCCCGGAAAAGGCCTGGAGTGGATGGGAATCATCTATCCTGGTGACTCTGATACC AGAT AC AGCCCGT CCTT CC AAGGCC AGGT C ACC AT CT C AGCCGAC AAGT CC AT C AGC ACCGCCTACCTGCAGTGGAGCAGCCTGAAGGCCTCGGACACCGCCGTGTATTACTGT GCGAGACCCAATGTTGACATAGTGGGAACTACCTACTTTGACTACTGGGGCCAGGG AACCCTGGTCACCGTCTCCTCA

3-4E SEQ ID NO: 17

GAGCTCGTGATGACTCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTC

ACCATCACTTGTCGGGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAG

AAGCCAGGGAAAGCCCCTAAGCTCCTGATCTATAAGGCGTCTAGTTTACAAAGTGG

GGTCCCATCAACGTTCAGCGGCAGTGGATCTGGGACAGAGTTCGCTCTCACCATCAG

CAGCCTGCAGCCTGATGATTTTGCAAGTTATTACTGCCAACAATATAATAGTTATCC

GTACACTTTTGGCCAGGGGACCAAGCTGGAGATCAAAGGTGGTTCCTCTAGATCTTC

CTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGCAGGTGCAGCTGGTGCAGTCTGG

GGCT GAGGT GAAGAAGCCTGGGGCCT C AGT GAAGGTTT CCTGC AAGGC AT CT GGAT

ACACCTTCACCAACTACTATATGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTTG

AGTGGAT GGGAAT AAT C AACCCT AGT GGT GGT GGC AC AAGTT AC AC AC AGAAGTTC

CAGGGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCT

GAGC AGCCT GAGAT CT GAGGAC ACGGCCGT GT ATT ACT GT GCGAGAGAT GGGTT GG

GCT ACT CTCT GGGC AGT GGCT GGT CT GACTACT GGGGCC AGGGAACCCT GGT C ACCG

TCTCCTCA

3-11E SEQ ID NO: 18

GAGCTCGTGATGACGCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTC

ACCATCACTTGTCGGGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAG

AAACCAGGGAAAGCCCCTAAACTCCTGATCTATAAGGCGTCTAGTTTACAAAGTGG

GGT CCC AT C AACGTT C AGCGGC AGT GGAT CT GGGAC AGAGTTC ACTCTC ACC ATC AG

C AGCCT GC AGCCT GAT GATTTT GC AACTT ATT ACTGCC AAC AATAT AAT AGTT AT CC

GTACACTTTTGGCCAGGGGACCAAGCTGGAGATCAAAGGTGGTTCCTCTAGATCTTC

CTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGGAGGTGCAGCTGGTGCAGTCTG

GGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTTTCCTGCAAGGCATCTGGA

TACACCTTCACCAACTACTATATGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTT GAGT GGAT GGGAAT AAT C AACCCTAGT GGT GGT GGC AC AAGTT AC AC AC AGAAGTT

CCAGGGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGC

TGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGCGAGGGATGGGTTG

GGCTACTCTCTGGGCAGTGGCTGGTCTGACTACTGGGGCCAGGGAACCCTGGTCACC

GTCTCCTCA

3-5E SEQ ID NO: 19

GAGCTCGAGCTGACTCAGCCACCCTCAGTGTCTGGGACCCCCGGGCAGAGGGTCAC

CAT CTCTT GTTCTGGAAGC AGCT CC AAC ATCGGGAGT AATTAT GT AT ACT GGT ACC A

GCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATAGGAATAATCAGCGGCCCTC

AGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCAT

CAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTATTGTGCAGCATGGGATGACA

GCCTGAGTGGCTGGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTAGGCGGTGGT

TCCTCTAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGCAGGTGCAG

CTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGC

AAGGCTTCTGGAGGCACCTTCAGCAGCTATGATACCAGCTGGGTGCGACAGGCCCC

TGGACAAGGGCTTGAGTGGATGGGAGGGATCATCCCTATGTTTGGTACACCAAAGT

ACGCACAGAAGTTCCAGGGCAGAGTCACCATTATCGCGGACGAATCCACGAACACA

GCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTTTATTATTGTGCG

AGAGCTCCTATAATAGTACTGGTATCCCGGGGTATGGACGTCTGGGGCCAAGGGAC

CACGGTCACCGTCTCCTCA

3-9E SEQ ID NO: 20

GAGCTCGAGCTGACTCAGCCACCCTCAGTGTCTGGGACCCCCGGGCAGAGGGTCAC

CATCTCTTGTTCTGGAAGCAGCTCCAACATCGGAACTAATTATGTATACTGGTACCA

GCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATAGGAGTAATCAGCGGCCCTC

AGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCAT

CAGTGGGCTCCAGTCTGAGGATGAGGCTGATTATTATTGTGCTGTCTGGGATGACAC

TCAACATCGTCGCGTGTTCGGCGGAGGGACCAAGGTGACCGTCCTAGGCGGTGGTTC

CTCTAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGCAGGTGCAGCT

GGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCA AGGCTT CT GGAGGC ACCTT C AGC AGCT AT GAT AT C AGCTGGGT GCGAC AGGCCCCTG

GAC AAGGGCTT GAGT GGAT GGGAGGGAT C ATCCCT ATCTTT GGT AC AGC AAACT AC

GCACAGAAGTTCCAGGGCAGAGTCACGATTACCGCGGACGAATCCACGAGCACAGC

CTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGCGA

GAGCTCCTATAATAGTACTGGTATCCCGGGGTATGGACGTCTGGGGCCAAGGGACC

ACGGTCACCGTCTCCTCA

3-38E SEQ ID NO: 21

GAGCTCGTGCTGACTCAGCCACCCTCAGCGTCTGGGACCCCCGGACAGAGGGTCAC

CATCTCTTGTTCTGGAAGCGGCTCCAACATCGGACGTAATTATGTATACTGGTACCA

GCAGCTCCCAGGAACGGCCCCCAAACTCTTCATCTTTAGGAATGATCAGCGGCCCTC

AGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCAT

CAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGCGCATCATGGGATGACA

GGC AGAGT GGTT GGGT GTTCGGCGGAGGC ACC AAGCT GACCGT CCT AGGCGGT GGT

TCCTCTAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGGAGGTGCAG

CTGGTGGAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTG

CAAGGCTTCTGGAGGCACCTTCAGCAGCTATGATATCAGCTGGGTGCGACAGGCCC

CTGGACAAGGGCTTGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGC AAGCT

ACGCACAGAAGTTCCAGGGCAGAGTCACGATTACCGCGGACGAATCCACGAGCACA

GCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTTTATTACTGTGCG

AGAGCTCCTATAATAGTACTGGTATCCCGGGGTATGGACGTCTGGGGCCAAGGGAC

CACGGTCACCGTCTCCTCG

3-48E SEQ ID NO: 22

GAGCTCGTGCTGACTCAGCCACCTTCAGCGTCTGGGGCCCCCGGGCAGAGGGTCAC

CATCTCTTGTTCTGGAAGCAGATCCAACATCGGAAGTAATTATGTATACTGGTACCA

GCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATAGGAATAATCAGCGGCCCTC

AGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCAT

CGGTGGACTCCGGTCCGACGATGAGGCTGATTATTACTGTGCAGCATGGGATGGCA

GTCTGCGTGCCTGGGTATTTGGCGGAGGCACCGAGCTGACCGTCCTCGGCGGTGGTT

CCTCTAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGCAGGTGCAGC TGGTGCAGTCTGGGGCTGAAGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGC

AAGGCTTCTGGAGGCTCCTTCAGTAGCTATGCTATCAACTGGGTGCGACAGGCCCCT

GGACAAGGGCTTGAGTGGATGGGAGGGATCATCCCTCCCTTTGGTGCAGCAGACTA

CGCACAGAAGTTCCAGGGCAGAATCACGATTACCGCGGACGATTCCGCGAGCACAG

CCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTTTATTACTGTGCGA

GAGCGCCTATAATAGTACTGGTATCCCGGGGTATGGACGTCTGGGGCCAAGGGACC

ACGGTCACCGTCTCCTCA

3-16E SEQ ID NO: 23

GAGCTCGTGCTGACTCAGCCACCTTCAGCGTCTGGGACCCCCGGGCAGAGGGTCAC

CATCTCTTGTTCTGGAAGCAGCTCCAACATCGGAAGGAATTATGTATACTGGTACCA

GCAACTCCCAGGAACGGCCCCCAAACTCCTCATCTATAAGAATAATCAGCGGCCCTC

AGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCAT

CAGTGGGCTCCAGTCTGAGGATGAGGCTGATTATTATTGTGCAGCATGGGGTGGCAG

CCT GAAGGGTT GGGT GTTCGGCGGAGGGACC AAGGT GACCGT CCT AGGT GGT GGTT

CCTCTAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGCAGGTGCAGC

TGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGC

AAGGCTTCCGGAGGCACCTTCAGCAACTTTGCTATCAGCTGGTTGCGACAGGCCCCT

GGACAAGGGCTTGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTA

CGCACAGAAGTTCCAGGGCAGAGTCACGATTACCGCGGACGAATCCACGAGCACAG

CCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGCG

ACCGGGCACAAAAACTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTC

A

3-29E SEQ ID NO: 24

GAGCTCGAGCTGACTCAGCCACCCTCAGTGTCTGGGACCCCCGGGCAGAGGGTCAC

CATCTCTTGTTCTGGAAGCAACTCCAACATCGGAAGTAATTATCTATACTGGTACCA

GCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATAGGAATAATCAGCGGCCCTC

AGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGACACCTCAGCCTCCCTGGCCAT

CAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGCAGCATGGGATGCCA

GCCTGAGTGGTTGGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTAGGCGGTGGT TCCTCTAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGCAGGTGCAG

CTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGC

AAGGCTTCTGGAGGCTCCTTCAGTAGCTATGCTATCAACTGGGTGCGACAGGCCCCT

GGACAAGGGCTTGAGTGGATGGGAGGGATCATCCCTCCCTTTGGTGCAGCAGACTA

CGCACAGAAGTTCCAGGGCAGAATCACGATTACCGCGGACGATTCCGCGAGCACAG

CCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGCG

AGAGGAGGGTCTAGCAGCAGCTGGCCACCAGTCGTTATGTACTACTGGGGCCAGGG

AACCCTGGTCACCGTTTCCTCA

3-34E SEQ ID NO: 25

GAGCTCGTGCTGACTCAGCCACCCTCAGCGTCTGGGACCCCCGGGCAGAGGGTCAC

CATCTCTTGTTCTGGAAGCAGCTCCAACATCGGAAGTAATTATGTATACTGGTACCA

GCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATAGGAATAATCAGCGGCCC

TCAGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCC

ATC AGT GGGCT CCGGT CCGAGGAT GAGGCT GAAT ATT ACT GT GC AGC AT GGGAT GA

CAGCCTGAGTGGTTATGTCTTCGGAACTGGGACCAAGGTGACCGTCCTAGGTGGTGG

TTCCTCTAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGGAGGTGCA

GCTGGTGCAGTCTGGAGGAGGCTTGATCCAGCCTGGGGGGTCCCTGAGACTCTCCTG

TGCAGCCTCTGGGTTCACCGTCAGTAGCAACTACATGACCTGGGTCCGCCAGGCTCC

AGGGAAGGGGCTGGAGTGGGTCTCATCCATTAGTAGTAGTAGTAGTTACATATACTA

CGC AGACT C AGTGAAGGGCCGATT C ACC ATCTCC AGAGAC AACGCC AAGAACT C AC

TGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCG

AGAC AT GAAAGGAACTT CT ACT AC AT GGACGT CT GGGGC A AAGGGACC ACGGT C AC

CGTCTCCTCA

3-37E SEQ ID NO: 26

GAGCTCGAGCTGACTCAGCCACCCTCAGTGTCTGGGACCCCCGGGCAGAGGGTCAC

CATCTCTTGTTCTGGAAGCAGCTCCAACATCGGAATTAATTATGTATACTGGTACCA

GCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATAGGAATAATCAGCGGCCCTC

AGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCAT

CAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGCAGCATGGGATGACA GCCTGAGTGGTTGGGTGTTCGGCGGAGGGACCAAGGTGACCGTCCTAGGCGGTGGT

TCCTCTAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGGAGGTGCAG

CTGGTGGAGTCTGGGGGAGGCCTGGTCAAGCCTGGGGGGTCCCTGAGACTCTCCTGT

GCAGCCTCTGGATTCACCTTCAGTCGCTATAGCATGAACTGGGTCCGCCAGGCTCCA

GGGAAGGGGCTGGAGTGGGTCTCATCCATTAGTAGTAGTAGTAGTTACATATACTAC

GCAGACTCAGTGAAGGGCCGATTCACCATCTCCAGAGACAACGCCAAGAACTCACT

GTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGG

GTAAACCGTATAGTAGCGGCTGGTCTGCGGAATACTGGGGCCAGGGAACCCTGGTC

ACCGTCCCCTCA

4-2E.230 SEQ ID NO: 27

GAGCTCGTGATGACCCAGTCTCCACTCTCCCTGCCCGTCACCCCTGGAGAGCCGGCC TCC AT CT CCTGC AGGT CT AGT C AGAGCCT CCTGC AT AGT GAT GGAT AC A ACT ATTT G GATTGGTACCTGCAGAAGCCAGGGCAGTCTCCACAGCTCCTGATCTATTTGGGTTCT TAT CGGGCCTCCGGGGT CCCT GAC AGGTTC AGT GGC AGT GGAT C AGGC AC AGATTTT AC ACTGAAAAT C AGC AGAGT GGAGGCT GAGGATGTT GGGGTTTATT ACT GC AT GCA AGCTCTACAAACTCCGTACACTTTTGGCCAGGGGACCAAGCTGGAGATCAAAGGTG GTTCCTCTAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGGAGGTGC AGCT GGT GC AGT CT GGGGCT GAGGT GAAGAAGCCT GGGT CCTCGGT GAAGGT CTCC TGCAAGGCTTCTGGAGGCACCTTCAGCAGCTATGCTATCAGCTGGGTGCGACAGGCC CCT GGAC AAGGGCTT GAGT GGAT GGGAGGGATC AT CCCT AT CTTT GGT AC AGC AAA CT ACGC AC AGA AGTT CCAGGGC AGAGT C ACGATT ACCGCGGACGAAT CC ACGAGC A CAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGT GCGAGAGAAGACTACTACTACTACGGTATGGACGTCTGGGGCCAAGGGACCACGGT CACCGTCTCCTCA

4-8E.230 SEQ ID NO: 28

GAGCTCACACTCACGCAGTCTCCAGGCACCCTGTCTTTGTCTCCAGGGGAAACAGCC

ACCCTCTCCTGTAGGGCCAGCCAGAGTGTTAGAAGCAACTACTTAGCCTGGTACCAG

CAAAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGGTGCATCCGGTAGGGCCACT

GGCATCCCAGACAGGTTCACTGGCAGCGGGTCTGGGACAGACTTCACTCTCACCATC AGCAGACTGGAGCCTGAAGATCTTGCAGTCTATTACTGTCAGCAGTATGGTCATGCA

CCTCGAATGTTCGGCCAAGGGACCAAGGTGGAAATCAAAGGTGGTTCCTCTAGATCT

TCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGCAGGTGCAGCTGGTGCAGTCT

GGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGTTTCTGG

AGGCACCTTCAACACCTATCCTTTCGCCTGGGTGCGACAGGCCCCTGGACAAGGGCC

TGAGTGGATGGGAGAAATCACCCCTCTCTTTGCCAGAGCAGACTACGCACAGAAGT

TCCAGGGCAGAGTCACGATTACCGCGGACGAATCCACGACCACAGTTTACATGGAA

CTGAACAGCCTGAGATCTGAGGACGCGGCCGTATATTACTGTACGACCCCTTTTACC

CACTCCTACTACTACGCCATGGACGCCTGGGGCCAAGGGACCACGGTCACCGTCTCC

CCA

2-6GL SEQ ID NO: 29

GAGCTCGTGCTGACTCAGCCACCCTCGGTGTCTGAAGCCCCCAGGCAGAGGGTCAC

CAT CCCCT GTT CT GGAGGC AGCTCC AAC ATCGGAAAT AAT GCT GT AAGCT GGT ACC A

GCAGCTCCCAGGAAAGGCTCCCAAACTCCTCATCTATTATGATGATCTCCTGCCCTC

AGGGGTCTCTGACCGATTCTCTGGCTCCAAGTCGGGCGCCTCTGCCTCCCTGGCCAT

CAGTGGGCTCCAGTCTGAGGATGAGGCTGATTATTACTGTGCAGCGTGGGATGACA

GCCTGAATGCTTGGGTGTTCGGCGGAGGGACCAAGGTGACCGTCCTAGGCGGTGGT

TCCTCTAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGCAGGTGCAG

CTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCGGTGAAGGTCTCCTG

CAAGGCTTCTGGATACACCTTCACCGGCTACTATCTCCACTGGCTGCGACAGGCCCC

TGGACAAGGGCCCGAGTGGATGGGATGGATCAACCCTAACAGTGGTGACACAAACT

TT GC AC AGAAGTTT C AGGGC AGT GT C ACCTTGACC AGGGAC ACGT CC AGC AAC AC A

GCCTACATGGAACTGAGCAGTCTGACATCTGGCGACACGGCCGTGTATTACTGTGCG

AGAGGGGGGTCGGGCCTGGGGGCTTTTGATGTTTGGGGCCAAGGGACAGTGGTCAC

CGTGTCTTCA

3-27GL SEQ ID NO: 30

GAGCTCGCCCTGACTCAGCCTCCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACC

ATCTCCTGCACTGGAACCAGCAGTGATGTTGGGAGTTATAACCTTGTCTCCTGGTAC

CAACAGCACCCAGGCAAAGCCCCCAAACTCATGATTTATGAGGGCAGTAAGCGGCC CTCAGGGGTCCCTGATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGAC

CATCTCGGGGCTCCAGGCTGAGGACGAGGCTCATTATTACTGCAGCTCATATACAAG

GAACAACGTGCTTTTCGGCGGAGGGACCAAGGTGACCGTCCTAGGTGGTGGTTCCTC

TAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGGAGGTGCAGCTGGT

GCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGG

GTTCTGGATACACCTTTACCAGCTACTGGATCACCTGGGTGCGCCAGGAGCCCGGGA

AAGGCCTGGAGTGGATGGGGATCATCTTTCCTGGTGACTCTGATACCAGATACAGCC

CGTCCTTCAAAGGCCAGGTCACCATCTCAGCCGACAAGTCCGTCAGCACCGCCTACC

TGCAGTGGAGCAGCCTGAAGGCCTCGGACACCGCCATCTATTATTGTGCGAGACTTG

GGTTAACTGGGGAGGACTCCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA

3-25GF SEQ ID NO: 31

GAGCTCGTGCTGACTCAGCCACCCTCAGCGTCTGGGACCCCCGGGCAGAGGGTCAC

CATCTCTTGTTCTGGAAGCAGCTCCAACATCGGAAGTAGTTATGTATACTGGTACCA

GCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATAGGAATAATCAGCGGCCCTC

AGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCTCTGGCCAT

CAGTGGGCTCCGGTCTGATGATGAGGCTCATTATTACTGTTCAGCCTGGGATCGCAG

CCTGAGTGAAGTGGTTTTCGGCGGAGGGACCCAGCTGACCGTCCTCGGCGGTGGTTC

CTCTAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGCAGGTGCAGCT

GCAGGAGTCGGGCCCACGACTGGTGAAGCCTTCGGAGACCCTGTCCCTCACCTGCA

GTGTCTCTGGATACTCCATCAGTGATGGTTATTACTGGGCCTGGGTCCGCCAGCCCC

C AGGGAAGGGGCT GGAGT GGAT CGGGACTATCC AT CGT GGT GGGAAT ACCT ACT AC

AACCCGTCCCTCAAGAGTCGACTCACCATATCTGCAGACACGTCCAAGAACCAGTTC

TCCCTGAACCTGAGGTCTGTGACCGCCGCAGACACGGCCGTGTATTACTGTGCGATG

GGGGGCGGGGGGCTACTATACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTC

A

3-29GF SEQ ID NO: 32

GAGCTCGAGCTGACTCAGCCACCCTCAGTGTCTGGGACCCCCGGGCAGAGGGTCAC

CATCTCTTGTTCTGGGAGCAGCTCCAACATCGGAAGTAATACTGTAAACTGGTACCA

GCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGATGATCTGCTGCCC TCAGGGGTCTCTGACCGATTCTCTGGCTCCAAGTCGGGCGCCTCTGCCTCCCTGGCC

ATCAGTGGGCTCCAGTCTGAGGATGAGGCTGATTATTACTGTGCAACATGGGATGAC

AGCCTGAATGCTTGGGTGTTCGGCGGAGGCACCAAGGTGACCGTCCTAGGCGGT

GGTTCCTCTAGATCTTCCTCCTCCGGTGGCTCGGGCGGTGGTGGGCAGGTGCAGCTG

GTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTCTCCTGCAA

GGCTTCTGGATACATGTTCACCGGCTACTATTTACACTGGGTGCGACAGGCCCCT

GGACAAGGGCTTGAGTGGATGGGATGGATCAACCCTCACAGTGGTGTCACATACTA

TGCACAGAAGTTTCAGGGCAGGGTCACCATGACCAGGGACACGTCCATCAGTACAG

CCTACGTGGAGCTGAGCAGACTGACATCTGGCGACACGGCCGTGTATTACTGTGCG

AGAGGGGGGTCGGGCCTGGGGGCTTTT GAT AT CTGGGGCC AAGGGAC AGT GGT C AC

CGTCTCTTCA

4-7E.230 SEQ ID NO: 65

GAGCTCGTGATGACCCAGTCTCCGGCCACCCTGTCTGCATCTGTGGGAGACAGAGTC

ACC ATC ACTT GCCGGGCC AGTC AGAGT AT AAGT AGAT GGTT AGCCT GGT AT C AGC AG

AAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGT

GGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACTATC

AGCAGCCTGCAGCCTGAAGATTTTGCAACTTACTATTGTCAACAGGCTAACAGTTTC

CCGATCACCTTCGGCCAAGGGACACGACTGGAGATTAAAGGTGGTTCCTCTAGA

TCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGGAGGTGCAGCTGTTGGAG

TCTGGGGGAGGTGTGGTACGGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCT

GGATTC ACCTTT GAT GATT ACGGC ATGAGCTGGGT CCGCC AAGCT CC AGGGAAG

GGGCT GGAGT GGGT CTCT GGT ATT AATT GGAAT GGTGGT AGC AC AGGTT AT GC AGAC

TCTGTGAAGGGCCGATTCACCATCTCCAGAGACAACGCCAAGAACTCACTGTATCTG

CAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGTCTA

TAGCAGGTGGCTTAGGTACTTTGAGTACTGGGGCCAGGGAACCCTGGTCACCGTCTC

CTCA

4-16E.230 SEQ ID NO: 66

GAGCTCGTGATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTC

ACCATCACTTGTCGGGCGAGTCAGGGTATTAGCAGCTGGTTAGCCTGGTATCAGCAG AAACCAGGGAAAGCCCCTAAACTCCTGATCTATGCTGCATCCAATTTACTAAGTGGG

GTCCCATCAAGGTTCAGCGGCAGTGGATATGGGACAGATTTCACTCTCACCATCAGC

AGCCTGCAGCCTGAAGATTTTGCA

ACTTACTATTGTCAACAGGCTAACAGTTTCCCCCTCACTTTCGGCGGAGGGACCAAG

CTGGAGATCAAAGGTGGTTCCTCTAAATCTTCCTCCTCTGGTGGCGGTGGCTCGGGC

GGTGGTGGGGAGGTGCAGCTGGTGCAGTCTGGGGGAGGTGTGGTACGGCCTGGG

GGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTTGATGATTATGGCATG

AGCTGGGTCCGCCAAGCTCCAGGGAAGGGGCTGGAGTGGGTCTCTGGTATTAATTG

GAATGGTGGTAGCACAGGTTATGCAGACTCTGTGAAGGGCCGATTCACCATCTCC

AGAGACAACGCCAAGAACTCCCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGA

CACGGCTGTGTATTACTGTGCGAGTATCTATAGCAGGTGGCTAAGGTACTTTGACTA

CTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA

3-22G SEQ ID NO: 67

GAGCTCGCCCTGACTCAGCCTCCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACC

ATCTCCTGCACTGGAACCAGCAGTGATGTTGTGCTTTATGACCTAGTCTCCTGGTAC C

AAC AAT ACCC AGGC AAAGCCCCC AAACTCTT GATTTTT GAT GTC AGT AAT CGG

CCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAGCACGGCCTCCCTG

ACC ATCTCT GGGCT CC AGGCT GAGGACGAGGCT GATTATT ACT GC AGCT CAT AT AC A

AGCAGCAGCACTAACTTATTCGGCGGAGGGACCAAGGTGACCGTCCTAGGCGGT

GGTTCCTCTAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGCAGATC

ACCTTGAAGGAGTCTGGTCCTGCGCTGGTGAAACCCACACAGACCCTCACGCTGACC

TGCACCTTCTCTGGGTTCTCACTCACCACTAGTGGAGTGGGTGTGGGCTGGATC

CGTCAGCCCCCAGGAAAGGCCCTGGAGTGGCTTGCACTCATTTATTGGAATGATGAT

AAGCGCTACAGCCCATCTCTGAAGAGCAGGCTCACCATCACCAAGGACACCTCCAA

AAACC AGGT GGT CCTT AC AAT GACC AAC AT GGACCCT GT GGAC AC AGCC AC AT AT

TACTGTGCACACAGGGACACCACGATTGGCAACTGGTTCGACCCCTGGGGCCAGGG

AACCCTGGTCACCGTCTCCTCA

3-14G SEQ ID NO: 33 ELTLTQSPGTLALSPGERATLSCRASQS VS SNYLAWYQQKPGQ APRLLIY GAS SRAPGIP DRF S GS GS GTDFTLTITSLQ AED V AVYY CQQ YFD SPLTF GGGTKVEIKGGS SRS S S S GGG GS GGGGQ V QL V Q S GAEVKKPGAS VKV S CKAS GYTF S S YDINWVRQ ATGQRPEWMGW MNPKSGDTGYAQTFQGRVTMTRSTSISTAYMELSSLRSEDTAMYYCTRGNRPIFDLFPF DLWGQGTLITV S S

3-17G SEQ ID NO: 34

ELVLTQPPSASGTPGQRVTISCSGSSSNIGLNFVYWYIQFPGTAPKLLIYRNNQRPSGVP D RISGSKSGTSASLAISGLRSEDEADYYCTSWDASLSAWVFGGGTKLTVLGGGSSRSSSSG GGGS GGGGQ V QLQES GPGLVKPSETLSLT CT V S GGS V S S GS YYW S WIRQPPGKGLEWIG SMF YSGRTYYNPSLN SRVTISGDTSKNQ V SLKLS S ATATDTAVYY CARRGSRLNTV GVP PAIELWGQGTLVTVSP

3-30G SEQ ID NO: 35

ELVLTQSPSASAAPGQRITISCSGSSSNIGINYVYWYQQVPGTAPKLLIYRNNQRPSGVP D

RFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLDGHWVFGGGTKVTVLGGGSSRS SS

SGGGGSGGGGQVQLQESGPGLVKPSETLSLTCTVSGGSISSGGYYWSWIRQPPGKGL EW IGSIF YSGRTYYNPSLN SRVTISGDTSKNQ V SLKLS SVTATDTAVYY CARRGTRLSETV AP AYEYWGQGTLVTVS S

3-4G SEQ ID NO: 36

ELVLTQSPSASGTPGQRVTISCSGSSSNIGSYFVYWYQQLPGTAPKLLIYRNSQRPSGVS D RFSGSKSGTSASLAISGLQSEDEADYYCASWDDSLSAWVFGGGTQLTVLGGGSSRSSSS GGGGS GGGGQ V QLQES GPGPVKPSETLSLT CT V S GGSIS SNT YYWEWIRQPPGKGLEWI GSMFYSGRTYYNPSLNSRVTISGDTSKNQVSLKLSSVTAADTAVYYCARRSLRYFDWSY DYWGQGTLVTVSS

3-29G SEQ ID NO: 37

ELGQTQQLSASGTPGQRVΉSCSGSSSNIGSHYVYWYQQLPGTAPKLLIYRNNQRPPGVP

DRFSGSKSATSASLAISGLQSEDEADYYCAAWDDSLDGHWVFGGGTQLTVLDGGSSR S

SSSGGGGSGGGGQVQLQESGPGLVKPSETLSLTCTVSGGSISSDTYYWDWIRQPPGR GL EWIGSINYSGNTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARLGSSGWY LPFD YWGQGTLVTVS S

3-3G SEQ ID NO: 38

EFEFTQPPSVSGTPGQRVTISCSGSSSSIGSNTVDWYQQVPGTAPKFFIYSNNQRPSGVP D RFSGSKSGTSASLAISGLQSEDEADYYCAAWDDSLNGYVLFGGGTKLTVLGGGSSRSSS SGGGGSGGGGQVQLVQSGAEVKKPGASVKVSCKASGYTFTDHYIHWVRQAPGQGLEW MGWINPNSGATNHAQRFQGRV SMTWDTS S STVNMELSRLRSDDTAVYY C ARGPYINY DYWGQGTLVTVSS

3-9G SEQ ID NO: 39

ELVLTQPPSASGTPGQRVTISCSGSSSNIGSNYVYWYQQLPGTAPKLLIYRNNQRPSGVP DRF S GSKS GTS ASL AI S GLRSEDE AD YY CGAWDGNLH AWVF GGGTKLTVLGGGS SRS S SSGGGGSGGGGQVQLVQSGTEVKKAGASVKVSCKASGNTFTSYYMHWVRQAPGQGL EWMGVIDPSGGSTTYAQKFQGGVTMTRDTSTSAVYMELSSLRSEDTAVYYCARGRLRG VIMSPFD YWGQGTLVTVS S

3-12G SEQ ID NO: 40

ELQMTQSPSTLSASVGDRVTITCRASQGIANWLAWYQQKPGKAPKLLIYAASSLQSGVP SRFSGSGSGTEFTLSISSLQPDDFATYYCQHYHSYPYTFGQGTKLEIKGGSSRSSSSGGG G S GGGGQ V QL V Q S GAEVKKPGAS VKV S CK AS GYTF SNYYMHWVRQ APGQGLEWMGII NPSGGGTRYAETFQGRITMTRDTSTNTVYMELSSLRSEDTAVYYCARDVEAYGSFMAY WGQGTLVTVSS

3-27G SEQ ID NO: 41

ELQMTQSPSTLSASIGDRVnACRASQSISSWLAWYQQKPGKAPKLLIYDASSLQSGVPS RF S GS GS GTDFTLTIS SLQPEDF AT YY CQQ S Y S VPLTF GGGTKVEIKGGS SRS S S S GGGGS GGGGQ V QL V Q S GAEVRKPGAS VKV S CE AS GYTLTNYYMHWVRQ APGQGLEWMGVIN PSGGSTNYAQKFQGRVTMTRDTSTSTVYMELSSLRPEDTAIYYCARDMRPNHYASGSF NYWGQGTLVTVSS 3-5G SEQ ID NO: 42

ELTLTQSPGTLSLSPGERATLSCRASQSVNNKYLAWYQQKPGQAPTLLIYGGSMRATGIP DRF S GS GSETDFTLTITRLEPEDF AVYYCQQ Y GRSPITFGQGTRLEIKGGS SRS S S S GGGG S GGGGQ V QF V Q S GAEVKKPGS S VKV S CKAS GGTF SNS AIS WVRQ APGQGFEWMGAIIPI FGTPNYAQRFQGRVTITADESTRTAYMEFGSFRSEDTAVYYCARDQPSWGSGWSPDHY YGFD VWGQGTTVTVS S

3-34G SEQ ID NO: 43

EFVFTQSPSASGTPGQRVTISCSGSSSNIGGNYVYWFQQFPGTAPKFFIYRNNQRPSGVP DRFSGSKSGTSASFAISGFQSEDEGDYYCAAWDASFKHRFFGGGTKVTVFGGGSSRSSS S GGGGS GGGGE V QFFES GGGF V QPGGSFRFS CS AS GFTF SNYWMS WVRQ APGKGFEW VS SIS S S SN YI Y Y AD 8MKOKRΉ SRDNAKNSFYFQMN SERAEDT AVYY C ARV Y SRWERY FEYWGQGTLVTVSS

4-4G.230 SEQ ID NO: 44

ELVLTQSPSSLSASVGDRVTIPCRASHNIKNYLNWYQQKPGKAPKLLIYAAYTLQGGVP SRFSGSGSGTDFM^SSLQPEDFATYYCQQSYSTPPAFGGGTKVDIKGGSSRSSSSGGGG SGGGGEVQLVESGGNLVQPGGSLRLSCSASGFTFEDYAMHWVRQAPGKGLEWVAGIF WNSDSIGYGDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARVGYDFWSGYSTP FDYWGQGTL V TV S S

4-5G.230 SEQ ID NO: 45

ELVLTQPPSVSGPPGQRAΉSCSGSSSNFGSY V WYQQLPGTAPKLLIHN NQRPSGVP DRF S GSKS GTS ASL AIS GLQ SEDE AE YY C AVWYD SLN GP VF GGGTQLTVLGGGS SRS S S S GGGGS GGGGQ V QLV Q S GAEVKKPGS S VKV S CKAS GGTFNT YPFTWVRQ APGQRLEWV GEITPLF GTAD YAQRFQGKV^S ADEPTTTVYMELNNLRSEDTAVYF CMTPFTHS YYY G MDAWGQGTTVTVS S

4-8G.230 SEQ ID NO: 46

ELTLTQSPGTLSLSPGERATLSCRASQNINSRLAWYQQKPGQAPRLLIYGASTRATGIPA R F S GS GS GTDFTLTISRLEPEDF AVYY CQH Y GS SPQLRTFGQGTKLEIKGGS SRS S S S GGGG S GGGGQ V QL V Q S GAEVKKPGS S VKV S CKAS GGTFNT YPFTWVRQ APGQRLEWV GEITP LF GTAD YAQRFQGRVTIS ADEPTTTVYMELNNLRSEDTAVYFCTTPFTFIS YYY GMD AW GQGTTVTVSS

3-47E SEQ ID NO: 47

ELELTQPPSVSGSPGQSVTISCTGTSSDVGGYKYVSWYQQHPGESPKLLIYEARKRPSGV PDRF S GSKS GKT ASLT V SDLQ ADDE A YYY C S S YGGSDNYLF GGGTKVTVLGGGS SRS S S SGGGGSGGGGEVQLVQSGAEVKKPGASVTVSCTASGYTFSSYDINWVRQATGQGPEW MGWMNPKSGETGYAQKFQGRVTMTRNTSIRTAYLELSSLRSDDTAIYYCARGNRPIFNL FPFDLWGQGTMVTVSS

3-3E SEQ ID NO: 48

ELELTQPPSVSGAPGQRVTISCTGSSSNIGAGYHVHWYQQFPGTAPKLLIYDNNNRPSGV

PDRFSGSKSGASASLAITGLQADDEADYYCQSYDSSLSGNWVFGGGTELTVLGGGSS RS

S S S GGGGS GGGGQ V QLV Q S GAE VKKPGESLKI S CKGS GYNF SNYWIGWVRQMPGKGLE

WMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAVYYCARPNYDI VGT

TYFDYWGQGTLVTVS

S

3-4E SEQ ID NO: 49

ELVMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYKASSLQSGVPS TFSGSGSGTEFALTISSLQPDDFASYYCQQYNSYPYTFGQGTKLEIKGGSSRSSSSGGGG S GGGGQ V QLV Q S GAEVKKPGAS VKV S CKAS GYTFTNYYMHWVRQ APGQGLEWMGIIN PSGGGTSYTQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDGLGYSLGSGWS DYWGQGTLVTVSS

3-11E SEQ ID NO: 50

ELVMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYKASSLQSGVPS TFSGSGSGTEFTLTISSLQPDDFATYYCQQYNSYPYTFGQGTKLEIKGGSSRSSSSGGGG S GGGGE V QLV Q S GAEVKKPGAS VKV S CKAS GYTFTNYYMHWVRQ APGQGLEWMGIINP SGGGTSYTQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDGLGYSLGSGWSD

YWGQGTLVTVSS

3-5E SEQ ID NO: 51

ELELTQPPSVSGTPGQRVTISCSGSSSNIGSNYVYWYQQLPGTAPKLLIYRNNQRPSGVP DRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLSGWVFGGGTKLTVLGGGSSRSSS S GGGGS GGGGQ V QL V Q S GAEVKKPGS S VKV S CKAS GGTF S S YDTS WVRQ APGQGLEW MGGIIPMF GTPKY AQKFQGRVTII ADES TNT A YMELS SLRSEDT AVYY C ARAPII VLV SR GMD VWGQGTTVTVS S

3-9E SEQ ID NO: 52

ELELTQPPSVSGTPGQRVTISCSGSSSNIGTNYVYWYQQLPGTAPKLLIYRSNQRPSGVP D RFSGSKSGTSASLAISGLQSEDEADYYCAVWDDTQHRRVFGGGTKVTVLGGGSSRSSSS GGGGS GGGGQ V QL V Q S GAEVKKPGS S VKV S CKAS GGTF S S YDI S WVRQ APGQGLEWM GGIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARAPIIVLVSRG M DVWGQGTTVTVSS

3-38E SEQ ID NO: 53

ELVLTQPPSASGTPGQRVTISCSGSGSNIGRNYVYWYQQLPGTAPKLFIFRNDQRPSGVP DRF S GSKS GTS ASL AIS GLRSEDE AD YY C AS WDDRQ S GWVFGGGTKLTVLGGGS SRS S S S GGGGS GGGGE V QLVES GAEVKKPGS S VKV S CKAS GGTF S S YDI S WVRQ APGQGLEW MGGIIPIF GTASYAQKFQGRVTITADESTSTAYMELS SLRSEDT AVYY C ARAPIIVLV SRG MD VWGQGTTVTVS S

3-48E SEQ ID NO: 54

ELVLTQPPSASGAPGQRVTISCSGSRSNIGSNYVYWYQQLPGTAPKLLIYRNNQRPSGVP DRFSGSKSGTS ASL AIGGLRSDDE AD YY C AAWDGSLRAWVF GGGTELTVLGGGS SRS S S S GGGGS GGGGQ V QL V Q S GAEVKKPGS S VKV S CKAS GGSF S S YAINWVRQ APGQGLEW MGGIIPPF GA AD Y AQKFQGRITIT ADD S AS T A YMELS SLRSED TAVY Y C ARAPIIVLV SRG MD VWGQGTTVTVS S 3-16E SEQ ID NO: 55

ELVLTQPPSASGTPGQRV^SCSGSSSNIGRNYVYWYQQLPGTAPKLLIYKNNQRPSGVP DRF S GSKS GTS ASL AI S GLQ SEDE AD Y Y C AAW GGSLKGWVFGGGTKVTVLGGGS SRS S S S GGGGS GGGGQ V QLV Q S GAEVKKPGS S VKV S CKAS GGTF SNF AIS WLRQ APGQGLEW MGGIIPIFGTANYAQKFQGRVΉTADESTSTAYMELSSLRSEDTAVYYCATGHKNFDYW GQGTLVTVSS

3-29E SEQ ID NO: 56

ELELTQPPSVSGTPGQRVTISCSGSNSNIGSNYLYWYQQLPGTAPKLLIYRNNQRPSGVP DRFSGSKSDTSASLAISGLRSEDEADYYCAAWDASLSGWVFGGGTKLTVLGGGSSRSSS S GGGGS GGGGQ V QLV Q S GAEVKKPGS S VKV S CKAS GGSF S S YAINWVRQ APGQGLEW MGGIIPPFGAAD YAQKFQGRITIT ADDS ASTAYMELS SLRSEDTAVYYC ARGGS S S S WPP VVMYYWGQGTLVTVS S

3-34E SEQ ID NO: 57

ELVLTQPPSASGTPGQRV^SCSGSSSNIGSNYVYWYQQLPGTAPKLLIYRNNQRPSGVP DRF S GSKS GTS ASL AI S GLRSEDE AEYY C AAWDD SLS GYVF GT GTKVTVLGGGS SRS S S S GGGGS GGGGE V QLV Q S GGGLIQPGGSLRLS C AAS GFT V S SNYMTWVRQ APGKGLEWV S SIS S S SSYIYYAD SVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARHERNFYYMD VWGKGTTVTVSS

3-37E SEQ ID NO: 58

ELELTQPPS VSGTPGQRVTISCS GS S SNIGINYVYWYQQLPGTAPKLLIYRNNQRPSGVPD RFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLSGWVFGGGTKVTVLGGGSSRSSSS GGGGS GGGGE V QLVES GGGLVKPGGSLRLS C AAS GFTF SRY SMNWVRQ APGKGLEWV S SIS S S S S YIYYADS VKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAGKPYS SGWS AE YWGQGTLVTVPS

4-2E.230 SEQ ID NO: 59

ELVMTQ SPLSLP VTPGEP ASI S CRS S Q SLLHSDGYNYLD WYLQKPGQ SPQLLI YLGS YRA S GVPDRF S GS GS GTDFTLKISRVE AED V GVYY CMQ ALQTP YTFGQGTKLEIKGGS SRS S S S GGGGS GGGGE V QL V Q S GAEVKKPGS S VKV S CK AS GGTF S S YAIS WVRQ APGQGLEW

MGGIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCAREDYYYY GM

DVWGQGTTVTVSS

4-8E.230 SEQ ID NO: 60

ELTLTQSPGTLSLSPGETATLSCRASQSVRSNYLAWYQQKPGQAPRLLIYGASGRATGIP DRFTGSGSGTDFTLTISRLEPEDLAVYYCQQYGHAPRMFGQGTKVEIKGGS SRS S SSGGG GS GGGGQ V QL V Q S GAEVKKPGS S VKV S CKV S GGTFNT YPF AWVRQ APGQGPEWMGEI TPLFARADYAQKFQGRVTITADESTTTVYMELNSLRSEDAAVYYCTTPFTHSYYYAMD AWGQGTTVTVSP

2-6GL SEQ ID NO: 61

ELVLTQPPS V SE APRQRVTIPCSGGS SNIGNNAV S WYQQLPGKAPKLLIYYDDLLPSGV S DRFSGSKSGASASLAISGLQSEDEADYYCAAWDDSLNAWVFGGGTKVTVLGGGSSRSS S S GGGGS GGGGQ V QLV Q S GAEVKKPGAS VKV S CKAS GYTFTGYYLHWLRQ APGQGPE WMGWINPNSGDTNFAQKFQGSVTLTRDTSSNTAYMELSSLTSGDTAVYYCARGGSGLG AFDVWGQGTVVTVSS

3-27GL SEQ ID NO: 62

ELALTQPPSVSGSPGQSITISCTGTSSDVGSYNLVSWYQQHPGKAPKLMIYEGSKRPSGV PDRF S GSKS GNT ASLTIS GLQ AEDE AHYYCS S YTRNNYLF GGGTKVTVLGGGS SRS S S S G GGGS GGGGE V QLV Q S GAEVKKPGESLRIS CKGS GYTFTS YWITWVRQEPGKGLEWMGII FPGDSDTRYSPSFKGQVΉSADKSVSTAYLQWSSLKASDTAIYYCARLGLTGEDSWGQG TLVTVSS

3-25GF SEQ ID NO: 63

ELVLTQPPSASGTPGQRVTISCSGSSSNIGSSYVYWYQQLPGTAPKLLIYRNNQRPSGVP D

RFSGSKSGTSASLAISGLRSDDEAHYYCSAWDRSLSEWFGGGTQLTVLGGGSSRSSS SG

GGGSGGGGQVQLQESGPRLVKPSETLSLTCSVSGYSISDGYYWAWVRQPPGKGLEWI G

TIHRGGNTYYNPSLKSRLTISADTSKNQFSLNLRSVTAADTAVYYCAMGGGGLLYYW G

QGTLVTVSS 3-29GF SEQ ID NO: 64

ELELTQPPSVSGTPGQIFV^SCSGSSSNIGSNTVNWYQQLPGTAPKLLIYYDDLLPSGVS D RFSGSKSGASASLAISGLQSEDEADYYCATWDDSLNAWVFGGGTKVTVLGGGSSRSSSS GGSGGGGQVQLVQSGAEVKKPGASVKVSCKASGYMFTGYYLHWVRQAPGQGLEWM GWINPHS GVTY Y AQKFQGRVTMTRDTSI S T A YVELSRLTS GD TAVY Y C ARGGS GLGAF DIWGQGTWTVSS

4-7E.230 SEQ ID NO: 68

ELVMTQ SP ATLS AS V GDRVTIT CRAS Q SISRWLAWY Q QKPGK APKLLI Y AAS SLQ S GVP SRF S GS GS GTDFTLTIS SLQPEDF AT YY CQQ AN SFPITF GQGTRLEIKGGS SRS S S S GGGGS GGGGE V QLLES GGGWRPGGSLRLS C AAS GFTFDD Y GMS WVRQ APGKGLEWV S GINW N GGS T GY AD S VKGRFTISRDNAKN SL YLQMN SLRAEDT AVYY C ARVY SRWLRYFEYW GQGTLVTVSS

4-16E.230 SEQ ID NO: 69

ELVMTQSPSSVSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKLLIYAASNLLSGVP SRFSGSGYGTDFTLTISSLQPEDFATYYCQQANSFPLTFGGGTKLEIKGGSSKSSSSGGG G S GGGGE V QL V Q S GGGWRPGGSLRLS C AAS GFTFDD Y GMS WVRQ APGKGLEWV S GIN

lnNOObTOUAObnKOEEΉbRONAKNbEUE MNbEEAEϋTAnUUEAbIUbElnEEUEϋU WGQGTLVTV S S

3-22G SEQ ID NO: 70

ELALTQPPSVSGSPGQSITISCTGTSSDWLYDLVSWYQQYPGKAPKLLIFDVSNRPSGVS NRFSGSKSGSTASLTISGLQAEDEADYYCSSYTSSSTNLFGGGTKVTVLGGGSSRSSSSG GGGSGGGGQITLKESGPALVKPTQTLTLTCTFSGFSLTTSGV GV GWIRQPPGKALEWLA LIYWNDDKRYSPSLKSRLTITKDTSKNQWLTMTNMDPVDTATYYCAHRDTTIGNWFD PWGQGTLVTV S S

Fusion Protein Sequences:

1005sqv Nucleotide SEQ ID NO: 71 CAACTTTGTATAGAAAAGTTGCTCGACATTGATTATTGACTAGTTATTAATAGTAAT

CAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTA

CGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATA

ATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTG

GAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGT

ACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTAC

ATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATT A

CCATGGTCGAGGTGAGCCCCACGTTCTGCTTCACTCTCCCCATCTCCCCCCCCTCCC C

ACCCCC AATTTT GT ATTT ATTT ATTTTTTA ATT ATTTT GT GC AGCGATGGGGGCGGGG

GGGGGGGGGGGGCGCGCGCCAGGCGGGGCGGGGCGGGGCGAGGGGCGGGGCGGG

GCGAGGCGGAGAGGTGCGGCGGCAGCCAATCAGAGCGGCGCGCTCCGAAAGTTTCC

TTTTATGGCGAGGCGGCGGCGGCGGCGGCCCTATAAAAAGCGAAGCGCGCGGCGGG

C GGG A GTC GC TGC GC GC TGC C TTC GC C C C GTGC C C C GC TC C GC C GC C GC C T C GC GC C

GCCCGCCCCGGCTCTGACTGACCGCGTTACTCCCACAGGTGAGCGGGCGGGACGGC

CCTTCTCCTCCGGGCTGTAATTAGCGCTTGGTTTAATGACGGCTTGTTTCTTTTCTG T

GGCTGCGTGAAAGCCTTGAGGGGCTCCGGGAGGGCCCTTTGTGCGGGGGGAGCGGC

TCGGGGGGTGCGTGCGTGTGTGTGTGCGTGGGGAGCGCCGCGTGCGGCTCCGCGCT

GCCCGGCGGCTGTGAGCGCTGCGGGCGCGGCGCGGGGCTTTGTGCGCTCCGCAGTG

TGCGCGAGGGGAGCGCGGCCGGGGGCGGTGCCCCGCGGTGCGGGGGGGGCTGCGA

GGGGAAC A AAGGCTGCGT GCGGGGT GT GT GCGT GGGGGGGTGAGC AGGGGGT GT G

GGCGCGTCGGTCGGGCTGCAACCCCCCCTGCACCCCCCTCCCCGAGTTGCTGAGCAC

GGCCCGGCTTCGGGTGCGGGGCTCCGTACGGGGCGTGGCGCGGGGCTCGCCGTGCC

GGGCGGGGGGTGGCGGCAGGTGGGGGTGCCGGGCGGGGCGGGGCCGCCTCGGGCC

GGGGAGGGCTCGGGGGAGGGGCGCGGCGGCCCCCGGAGCGCCGGCGGCTGTCGAG

GCGCGGCGAGCCGC AGCC ATT GCCTTTT AT GGT AAT CGT GCGAGAGGGCGC AGGGA

CTTCCTTTGTCCCAAATCTGTGCGGAGCCGAAATCTGGGAGGCGCCGCCGCACCCCC

TCTAGCGGGCGCGGGGCGAAGCGGTGCGGCGCCGGCAGGAAGGAAATGGGCGGGG

AGGGCCTTCGTGCGTCGCCGCGCCGCCGTCCCCTTCTCCCTCTCCAGCCTCGGGGCT

GTCCGCGGGGGGACGGCTGCCTTCGGGGGGGACGGGGCAGGGCGGGGTTCGGCTTC

TGGCGTGTGACCGGCGGCTCTAGAGCCTCTGCTAACCATGTTCATGCCTTCTTCTTT T

TCCTACAGCTCCTGGGCAACGTGCTGGTTATTGTGCTGTCTCATCATTTTGGCAAAG A ATT GC AAGTTT GT AC AAAA AAGC AGGCTGCC ACC ATGGAGAC AGAC AC ACT CCTGC

TATGGGTACTGCTGCTCTGGGTTCCAGGTTCCACTGGTGACACCAGCGGCGACGACG

ACGACAAGGCCAAGAAGAAGCTGCCCAAGTGCCAGAAGCAGGAGGACTGCGGCAG

CT GGGACCTGAAGT GC AAC AACGT GACC AAGA AGT GCGAGT GC AGGAACC AGGT GT

GCGGCAGGGGCTGCCCCAAGGAGAGGTACCAGAGGGACAAGTACGGCTGCAGGAA

GTGCCT GT GC AAGGGCT GCGACGGCTT C AAGT GC AGGCT GGGCT GC ACCT ACGGCTT

CAAGACCGACAAGAAGGGCTGCGAGGCCTTCTGCACCTGCAACACCAAGGAGACCG

CCTGCGTGAACATCTGGTGCACCGACCCCTACAAGTGCAACCCCGAGAGCGGCAGG

TGCGAGGACCCCAACGAGGAGTACGAGTACGACTACGAGGGTGGTTCCTCTAGATC

TTCCTCCTCTGGTGGCGGTGGCTCGGGCGGTGGTGGGGAGCTCACACTCACGCAGTC

TCCAGGCACCCTGGCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAG

TCAGAGTGTTAGCAGCAACTACTTAGCCTGGTACCAGCAGAAACCTGGCCAGGCTC

CC AGGCTCCT CAT CT AT GGT GC AT CC AGC AGGGCCCCT GGC AT CCC AGAC AGGTT C A

GTGGCAGTGGGTCTGGGACAGATTTCACTCTCACCATCACCAGCCTGCAGGCTGAAG

ATGTGGCAGTTTATTACTGTCAGCAATATTTTGATAGTCCGCTCACTTTCGGCGGAG

GGACCAAGGTGGAAATCAAAGGTGGTTCCTCTAGATCTTCCTCCTCTGGTGGCGGTG

GCTCGGGCGGTGGTGGGCAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAG

CCTGGGGCCTCAGTGAAGGTCTCCTGCAAGGCTTCTGGATACACCTTCAGCAGTTAT

GACATCAATTGGGTGCGACAGGCCACTGGACAACGGCCTGAGTGGATGGGGTGGAT

GAACCCTAAAAGTGGTGACACAGGCTACGCACAGACCTTCCAGGGCAGAGTCACCA

TGACCAGGAGCACCTCCATAAGTACAGCCTATATGGAACTGAGCAGCCTGAGATCT

GAAGACACGGCCATGTATTATTGTACGAGAGGCAACCGACCGATTTTTGATTTATTT

CCTTTTGATCTCTGGGGCCAAGGGACACTGATCACCGTCTCTTCAGCTTCCACCAAG

GGCCCATCGGTCACTAGTGGCCAGGCCGGCCAGCACCATCACCATCACCATGGCGC

ATACCCGTACGACGTTCCGGACTACGCTTCTTAAACCCAGCTTTCTTGTACAAAGTG

GTGATGGCCGGCCGCTTCGAGCAGACATGATAAGATACATTGATGAGTTTGGACAA

ACCACAACTAGAATGCAGTGAAAAAAATGCTTTATTTGTGAAATTTGTGATGCTATT

GCTTT ATTT GTAACC ATT ATA AGCT GC AAT AAAC AAGTT AAC AAC AAC AATT GC ATT

C ATTTT AT GTTT C AGGTT C AGGGGGAGGT GTGGGAGGTTTTTTAAAGC AAGT AAAAC

CTCTACAAATGTGGTAGCGGCCGCGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGC

TGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATAC GGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCA

GC AAAAGGCC AGGAACCGT AAAAAGGCCGCGTT GCT GGCGTTTTT CC AT AGGCT CC

GCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCG

ACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCT

GTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCTCTTCGGGAAGCGTG G

CGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCA A

GCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAA

CTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCAC

TGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGT

GGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGA

AGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACC

GCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGG

ATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAA

CTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCT

TTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTC

TGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCG T

TCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTA

CCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGAT

TT AT C AGC AAT AAACC AGCC AGCCGGAAGGGCCGAGCGC AGAAGT GGT CCT GC AAC

TTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTC

GCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACG

CTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTAC

ATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGT

CAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTC

TCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAA G

TCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGG

GATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCT

TCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCC

ACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGA G

CAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATG

TTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTG T CT CAT GAGCGGAT AC AT ATTT GAAT GT ATTTAGAA AAAT AAAC AAAT AGGGGTT CCG

CGCACATTTCCCCGAAAAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACA

TTAACCTATAAAAATAGGCGTATCACGAGGCCCTTTCGTCGGCGCGCCGCGGCCGC

1012qxm Nucleotide SEQ P) NO: 72

CAACTTTGTATAGAAAAGTTGCTCGACATTGATTATTGACTAGTTATTAATAGTAAT

CAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTA

CGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATA

ATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTG

GAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGT

ACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTAC

ATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATT A

CCATGGTCGAGGTGAGCCCCACGTTCTGCTTCACTCTCCCCATCTCCCCCCCCTCCC C

ACCCCC AATTTT GT ATTT ATTT ATTTTTTA ATT ATTTT GT GC AGCGATGGGGGCGGGG

GGGGGGGGGGGGCGCGCGCCAGGCGGGGCGGGGCGGGGCGAGGGGCGGGGCGGG

GCGAGGCGGAGAGGTGCGGCGGCAGCCAATCAGAGCGGCGCGCTCCGAAAGTTTCC

TTTTATGGCGAGGCGGCGGCGGCGGCGGCCCTATAAAAAGCGAAGCGCGCGGCGGG

C GGG A GTC GC TGC GC GC TGC C TTC GC C C C GTGC C C C GC TC C GC C GC C GC C T C GC GC C

GCCCGCCCCGGCTCTGACTGACCGCGTTACTCCCACAGGTGAGCGGGCGGGACGGC

CCTTCTCCTCCGGGCTGTAATTAGCGCTTGGTTTAATGACGGCTTGTTTCTTTTCTG T

GGCTGCGTGAAAGCCTTGAGGGGCTCCGGGAGGGCCCTTTGTGCGGGGGGAGCGGC

TCGGGGGGTGCGTGCGTGTGTGTGTGCGTGGGGAGCGCCGCGTGCGGCTCCGCGCT

GCCCGGCGGCTGTGAGCGCTGCGGGCGCGGCGCGGGGCTTTGTGCGCTCCGCAGTG

TGCGCGAGGGGAGCGCGGCCGGGGGCGGTGCCCCGCGGTGCGGGGGGGGCTGCGA

GGGGAAC A AAGGCTGCGT GCGGGGT GT GT GCGT GGGGGGGTGAGC AGGGGGT GT G

GGCGCGTCGGTCGGGCTGCAACCCCCCCTGCACCCCCCTCCCCGAGTTGCTGAGCAC

GGCCCGGCTTCGGGTGCGGGGCTCCGTACGGGGCGTGGCGCGGGGCTCGCCGTGCC

GGGCGGGGGGTGGCGGCAGGTGGGGGTGCCGGGCGGGGCGGGGCCGCCTCGGGCC

GGGGAGGGCTCGGGGGAGGGGCGCGGCGGCCCCCGGAGCGCCGGCGGCTGTCGAG

GCGCGGCGAGCCGC AGCC ATT GCCTTTT AT GGT AAT CGT GCGAGAGGGCGC AGGGA

CTTCCTTTGTCCCAAATCTGTGCGGAGCCGAAATCTGGGAGGCGCCGCCGCACCCCC TCTAGCGGGCGCGGGGCGAAGCGGTGCGGCGCCGGCAGGAAGGAAATGGGCGGGG

AGGGCCTTCGTGCGTCGCCGCGCCGCCGTCCCCTTCTCCCTCTCCAGCCTCGGGGCT

GTCCGCGGGGGGACGGCTGCCTTCGGGGGGGACGGGGCAGGGCGGGGTTCGGCTTC

TGGCGTGTGACCGGCGGCTCTAGAGCCTCTGCTAACCATGTTCATGCCTTCTTCTTT T

TCCTACAGCTCCTGGGCAACGTGCTGGTTATTGTGCTGTCTCATCATTTTGGCAAAG A

ATT GC AAGTTT GT AC AAAA AAGC AGGCTGCC ACC ATGGAGAC AGAC AC ACT CCTGC

TATGGGTACTGCTGCTCTGGGTTCCAGGTTCCACTGGTGACGGTGGTTCCTCTAGAT C

TTCCGAGAACCTGTACTTCCAGGGCGCCGCCGAGCTCACACTCACGCAGTCTCCAGG

CACCCTGGCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAG

TGTTAGCAGCAACTACTTAGCCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCT

CCTCATCTATGGTGCATCCAGCAGGGCCCCTGGCATCCCAGACAGGTTCAGTGGCAG

TGGGTCTGGGACAGATTTCACTCTCACCATCACCAGCCTGCAGGCTGAAGATGTGGC

AGTTTATTACTGTCAGCAATATTTTGATAGTCCGCTCACTTTCGGCGGAGGGACCAA

GGTGGAAATCAAAGGTGGTTCCTCTAGATCTTCCTCCTCTGGTGGCGGTGGCTCGGG

CGGTGGTGGGCAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGG

CCTCAGTGAAGGTCTCCTGCAAGGCTTCTGGATACACCTTCAGCAGTTATGACATCA

ATTGGGTGCGACAGGCCACTGGACAACGGCCTGAGTGGATGGGGTGGATGAACCCT

AAAAGTGGTGACACAGGCTACGCACAGACCTTCCAGGGCAGAGTCACCATGACCAG

GAGCACCTCCATAAGTACAGCCTATATGGAACTGAGCAGCCTGAGATCTGAAGACA

CGGCCATGTATTATTGTACGAGAGGCAACCGACCGATTTTTGATTTATTTCCTTTTG A

TCTCTGGGGCCAAGGGACACTGATCACCGTCTCTTCAGCTTCCACCAAGGGCCCATC

GGTCACTAGTGGCCAGGCCGGCCAGCACCATCACCATCACCATGGCGCATACCCGT

ACGACGTTCCGGACTACGCTTCTGGTGGTTCCTCTAGATCTTCCTCCTCTGGTGGCG G

TGGCTCGGGCGGTGGTGGGCTGGCCGAGGCCGCCGCCAAGGAGGCCGCCGCCAAGG

A GGC C GC C GC C A A GG A GGC C GC C GC C A A GG A GGC C GC C GC C A A GGC C GC C GC C GG

TGGTTCCTCTAGATCTTCCACCAGCGGCGACGACGACGACAAGGCCAAGAAGAAGC

TGCCCAAGTGCCAGAAGCAGGAGGACTGCGGCAGCTGGGACCTGAAGTGCAACAAC

GTGACCAAGAAGTGCGAGTGCAGGAACCAGGTGTGCGGCAGGGGCTGCCCCAAGG

AGAGGTACCAGAGGGACAAGTACGGCTGCAGGAAGTGCCTGTGCAAGGGCTGCGAC

GGCTT C AAGT GC AGGCT GGGCT GC ACCT ACGGCTT C AAGACCGAC AAGAAGGGCT G

CGAGGCCTTCTGCACCTGCAACACCAAGGAGACCGCCTGCGTGAACATCTGGTGCA CCGACCCCTACAAGTGCAACCCCGAGAGCGGCAGGTGCGAGGACCCCAACGAGGA

GTACGAGTACGACTACGAGGGTGGTTCCTCTAGATCTTCCTAAACCCAGCTTTCTTG

TACAAAGTGGTGATGGCCGGCCGCTTCGAGCAGACATGATAAGATACATTGATGAG

TTT GGAC AA ACC AC AACT AGA ATGC AGT GAAAAA AATGCTTT ATTTGTGAAATTT GT

GATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATAAACAAGTTAACAACAAC

AATT GC ATT C ATTTT ATGTTT C AGGTT C AGGGGGAGGT GT GGGAGGTTTTTT AAAGC

AAGTAAAACCTCTACAAATGTGGTAGCGGCCGCGGCGCTCTTCCGCTTCCTCGCTCA

CTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGG

CGGT AAT ACGGTT AT CC AC AGAAT C AGGGGAT AACGC AGGAAAGA AC AT GT GAGC A

AAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCA

TAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGC

GAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTG

CGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCTCTTCG G

GAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCG

TTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCT

TATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGG

C AGC AGCC ACT GGT AAC AGGATT AGC AGAGCGAGGT AT GT AGGCGGTGCTAC AGAG

TTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGC

GCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAA

C AAACC ACCGCT GGT AGCGGT GGTTTTTTT GTTT GC A AGC AGC AGATTACGCGC AGA

AAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGG

AACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACC

TAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAA

ACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGT

CTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGG G

AGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCG

GCT CC AGATTT AT C AGC AAT AAACC AGCC AGCCGGAAGGGCCGAGCGCAGAAGT GG

TCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGT A

AGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTG

GTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGG

CGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCG ATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTG CATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACT CAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGT C AAT ACGGGAT AAT ACCGCGCC AC AT AGC AGA ACTTTAAAAGTGCTC AT C ATT GGA AAACGTT CTT CGGGGCGAAAACT CT C AAGGAT CTT ACCGCT GTTGAGAT CC AGTT CG ATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTT CT GGGT GAGC AAAAAC AGGAAGGC AAAATGCCGC AAAAAAGGGAAT AAGGGCGAC ACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAG GGTT ATT GT CT CAT GAGCGGAT AC AT ATTT GAAT GT ATTT AGAAAAAT AAAC AAAT A GGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTCTAAGAAACCATTATT ATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCCCTTTCGTCGGCGCGCC GCGGCCGC

1048nvw Nucleotide SEQ ID NO: 73

TAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCCCCTCTAGAAATAA

TTTTGTTTAACTTTAAGAAGGAGATATACCATGAAAAAAACCGCGATTGCGATTGCG

GTGGCGCTGGCGGGCTTTGCGACCGTGGCGCAGGCGGCGGCGAGCAGCAGCCGCTG

C A C C T A TG A TC A TTGGTGC A GC C A TGGC GGC A GC A GC C GC A GC A GC A GC A GC GGC G

GCGGCGGCAGCGGCGGCGGCGGCGAACTGACCCTGACCCAGAGCCCGGGCACCCTG

GCGCTGAGCCCGGGCGAACGCGCGACCCTGAGCTGCCGCGCGAGCCAGAGCGTGAG

CAGCAACTATCTGGCGTGGTATCAGCAGAAACCGGGCCAGGCGCCGCGCCTGCTGA

TTTATGGCGCGAGCAGCCGCGCGCCGGGCATTCCGGATCGCTTTAGCGGCAGCGGC

AGCGGCACCGATTTTACCCTGACCATTACCAGCCTGCAGGCGGAAGATGTGGCGGT

GTATTATTGCCAGCAGTATTTTGATAGCCCGCTGACCTTTGGCGGCGGCACCAAAGT

GGAAATTAAAGGCGGCAGCAGCCGCAGCAGCAGCAGCGGCGGCGGCGGCAGCGGC

GGCGGCGGCCAGGTGCAGCTGGTGCAGAGCGGCGCGGAAGTGAAAAAACCGGGCG

CGAGCGTGAAAGTGAGCTGCAAAGCGAGCGGCTATACCTTTAGCAGCTATGATATT

AACTGGGTGCGCCAGGCGACCGGCCAGCGCCCGGAATGGATGGGCTGGATGAACCC

GAAAAGCGGCGATACCGGCTATGCGCAGACCTTTCAGGGCCGCGTGACCATGACCC

GCAGCACCAGCATTAGCACCGCGTATATGGAACTGAGCAGCCTGCGCAGCGAAGAT

ACCGCGATGTATTATTGCACCCGCGGCAACCGCCCGATTTTTGATCTGTTTCCGTTT G ATCTGTGGGGCCAGGGCACCCTGATTACCGTGAGCAGCGCGAGCACCAAAGGCCCG

AGCGTGACCAGCGGCCAGGCGGGCCAGCATCATCATCATCATCATGGCGCGTATCC

GTATGATGTGCCGGATTATGCGAGCTAAGGATCCGCTGCTAACAAAGCCCGAAAGG

AAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGGCCT

CTAAACGGGTCTTGAGGGGTTTTTTGCTGAAAGGAGGAACTATATCCGGATATCCCG

CAAGAGGCCCGGCAGTACCGGCATAACCAAGCCTATGCCTACAGCATCCAGGGTGA

CGGTGCCGAGGATGACGATGAGCGCATTGTTAGATTTCATACACGGTGCCTGACTGC

GTTAGCAATTTAACTGTGATAAACTACCGCATTAAAGCTTATCGATGATAAGCTGTC

AAAC AT GAGA ATT CTT GAAGACGAA AGGGCCTCGT GAT ACGCCT ATTTTT ATAGGTT

AATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTG

CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATG A

GAC AAT AACCCTGAT AAAT GCTT C AAT AAT ATT GAAAAAGGAAGAGT AT GAGT ATT

CAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTT GC

TC ACCC AGAAACGCT GGT GAAAGT AAAAGAT GCT GAAGAT C AGTT GGGT GC ACGAG

TGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCG

AAGAACGTTTTCC AAT GAT GAGC ACTTTT AAAGTT CT GCT AT GT GGCGCGGT ATT AT

CCCGTGTTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATG

ACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTA

AGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTT

CT GAC A ACGAT CGGAGGACCGAAGGAGCT AACCGCTTTTTTGC AC AAC AT GGGGGA

TCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACG

ACGAGCGTGACACCACGATGCCTGCAGCAATGGCAACAACGTTGCGCAAACTATTA

ACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCG

GATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCT

GATAAATCTGGAGCCGGTGAGCGTGGGTCACGCGGTATCATTGCAGCACTGGGGCC

AGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTAT

GGAT GAACGAAAT AGAC AGAT CGCTGAGAT AGGT GCCT C ACTGATTA AGC ATT GGT

AACT GT C AGACC AAGTTT ACT CAT AT AT ACTTT AGATT GATTT AAA ACTTC ATTTTTA

ATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTA

ACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTC

TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCT ACC AGCGGT GGTTTGTTT GCCGGAT C AAGAGCT ACC AACT CTTTTT CCGAAGGT AAC

TGGCTTC AGC AGAGCGC AGAT ACC AAAT ACT GT CCTTCT AGT GT AGCCGT AGTT AGG

CCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTT

ACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACG

ATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGC

CCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGA

GAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCA

GGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTT

TATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCG T

CAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTG

GCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTG GA

TAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGA

GCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGGGCGCCTGATGCGGTATTTTCTCC

TT ACGC AT CT GT GCGGT ATTT C AC ACCGC AAT GGT GC ACT CT C AGT AC AATCTGCT CT

GATGCCGCATAGTTAAGCCAGTATACACTCCGCTATCGCTACGTGACTGGGTCATGG

CTGCGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCC

CGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGT

TTTCACCGTCATCACCGAAACGCGCGAGGCAGCTGCGGTAAAGCTCATCAGCGTGG

TCGTGAAGCGATTCACAGATGTCTGCCTGTTCATCCGCGTCCAGCTCGTTGAGTTTC T

CC AGAAGCGTT AAT GTCTGGCTT CT GAT AAAGCGGGCC ATGTT AAGGGCGGTTTTTT

CCTGTTTGGTCACTGATGCCTCCGTGTAAGGGGGATTTCTGTTCATGGGGGTAATGA

TACCGATGAAACGAGAGAGGATGCTCACGATACGGGTTACTGATGATGAACATGCC

CGGTTACTGGAACGTTGTGAGGGTAAACAACTGGCGGTATGGATGCGGCGGGACCA

GAGAAAAATCACTCAGGGTCAATGCCAGCGCTTCGTTAATACAGATGTAGGTGTTCC

ACAGGGTAGCCAGCAGCATCCTGCGATGCAGATCCGGAACATAATGGTGCAGGGCG

CTGACTTCCGCGTTTCCAGACTTTACGAAACACGGAAACCGAAGACCATTCATGTTG

TTGCTCAGGTCGCAGACGTTTTGCAGCAGCAGTCGCTTCACGTTCGCTCGCGTATCG

GTGATTCATTCTGCTAACCAGTAAGGCAACCCCGCCAGCCTAGCCGGGTCCTCAACG

ACAGGAGCACGATCATGCGCACCCGTGGCCAGGACCCAACGCTGCCCGAGATGCGC

CGCGTGCGGCTGCTGGAGATGGCGGACGCGATGGATATGTTCTGCCAAGGGTTGGTT

TGCGCATTCACAGTTCTCCGCAAGAATTGATTGGCTCCAATTCTTGGAGTGGTGAAT CCGTTAGCGAGGTGCCGCCGGCTTCCATTCAGGTCGAGGTGGCCCGGCTCCATGCAC

CGCGACGC AACGCGGGGAGGC AGAC AAGGT AT AGGGCGGCGCCT AC AAT CC AT GCC

AACCCGTTCCATGTGCTCGCCGAGGCGGCATAAATCGCCGTGACGATCAGCGGTCC

AATGATCGAAGTTAGGCTGGTAAGAGCCGCGAGCGATCCTTGAAGCTGTCCCTGAT

GGTCGTCATCTACCTGCCTGGACAGCATGGCCTGCAACGCGGGCATCCCGATGCCGC

CGGAAGCGAGAAGAATCATAATGGGGAAGGCCATCCAGCCTCGCGTCGCGAACGCC

AGCAAGACGTAGCCCAGCGCGTCGGCCGCCATGCCGGCGATAATGGCCTGCTTCTC

GCCGAAACGTTTGGTGGCGGGACCAGTGACGAAGGCTTGAGCGAGGGCGTGCAAGA

TTCCGAATACCGCAAGCGACAGGCCGATCATCGTCGCGCTCCAGCGAAAGCGGTCC

TCGCCGAAAATGACCCAGAGCGCTGCCGGCACCTGTCCTACGAGTTGCATGATAAA

GAAGACAGTCATAAGTGCGGCGACGATAGTCATGCCCCGCGCCCACCGGAAGGAGC

TGACTGGGTTGAAGGCTCTCAAGGGCATCGGTCGAGATCCCGGTGCCTAATGAGTG

AGCTAACTTACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTG

TCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATT

GGGCGCCAGGGTGGTTTTTCTTTTCACCAGTGAGACGGGCAACAGCTGATTGCCCTT

CACCGCCTGGCCCTGAGAGAGTTGCAGCAAGCGGTCCACGCTGGTTTGCCCCAGCA

GGCGAAAATCCTGTTTGATGGTGGTTAACGGCGGGATATAACATGAGCTGTCTTCGG

TATCGTCGTATCCCACTACCGAGATATCCGCACCAACGCGCAGCCCGGACTCGGTAA

TGGCGCGCATTGCGCCCAGCGCCATCTGATCGTTGGCAACCAGCATCGCAGTGGGA

ACGATGCCCTCATTCAGCATTTGCATGGTTTGTTGAAAACCGGACATGGCACTCCAG

TCGCCTTCCCGTTCCGCTATCGGCTGAATTTGATTGCGAGTGAGATATTTATGCCAG C

CAGCCAGACGCAGACGCGCCGAGACAGAACTTAATGGGCCCGCTAACAGCGCGATT

TGCTGGTGACCCAATGCGACCAGATGCTCCACGCCCAGTCGCGTACCGTCTTCATGG

GAGAAAATAATACTGTTGATGGGTGTCTGGTCAGAGACATCAAGAAATAACGCCGG

AACATTAGTGCAGGCAGCTTCCACAGCAATGGCATCCTGGTCATCCAGCGGATAGTT

AATGATCAGCCCACTGACGCGTTGCGCGAGAAGATTGTGCACCGCCGCTTTACAGG

CTTCGACGCCGCTTCGTTCTACCATCGACACCACCACGCTGGCACCCAGTTGATCGG

CGCGAGATTTAATCGCCGCGACAATTTGCGACGGCGCGTGCAGGGCCAGACTGGAG

GTGGCAACGCCAATCAGCAACGACTGTTTGCCCGCCAGTTGTTGTGCCACGCGGTTG

GGAATGTAATTCAGCTCCGCCATCGCCGCTTCCACTTTTTCCCGCGTTTTCGCAGAA A

CGTGGCTGGCCTGGTTCACCACGCGGGAAACGGTCTGATAAGAGACACCGGCATAC TCTGCGACATCGTATAACGTTACTGGTTTCACATTCACCACCCTGAATTGACTCTCTT

CCGGGCGCTATCATGCCATACCGCGAAAGGTTTTGCGCCATTCGATGGTGTCCGGGA

TCTCGACGCTCTCCCTTATGCGACTCCTGCATTAGGAAGCAGCCCAGTAGTAGGTTG

AGGCCGTTGAGCACCGCCGCCGCAAGGAATGGTGCATGCAAGGAGATGGCGCCCAA

CAGTCCCCCGGCCACGGGGCCTGCCACCATACCCACGCCGAAACAAGCGCTCATGA

GCCCGAAGTGGCGAGCCCGATCTTCCCCATCGGTGATGTCGGCGATATAGGCGCCA

GCAACCGCACCTGTGGCGCCGGTGATGCCGGCCACGATGCGTCCGGCGTAGAGGAT

CGAGATCTCGATCCCGCGAAAT

1005sqv Amino Acid SEQ P ⁾ NO: 74

METDTLLLWVLLLWVPGSTGDTSGDDDDKAKKKLPKCQKQEDCGSWDLKCNNYTKK CECRNQ V CGRGCPKERY QRDKY GCRKCLCKGCDGFKCRLGCTY GFKTDKKGCE AFCT CNTKETAC VNTWCTDP YKCNPES GRCEDPNEE YE YD YEGGS SRS S S S GGGGS GGGGELT LTQ SPGTL ALSPGERATLS CRAS Q S VS SN YLAWYQQKPGQ APRLLI Y GAS SRAPGIPDRF S GS GS GTDFTLTITSLQ AED V AVYY CQ Q YFD SPLTF GGGTKVEIKGGS SRS S S S GGGGS G GGGQ V QL V Q S GAEVKKPGAS VKV S CKAS GYTF S S YDINWVRQ ATGQRPEWMGWMNP KSGDTGYAQTFQGRVTMTRSTSISTAYMELSSLRSEDTAMYYCTRGNRPIFDLFPFDLW GQGTLITVSSASTKGPSVTSGQAGQHHHHHHGAYPYDVPDYAS

1012qxm Amino Acid SEQ P) NO: 75

METDTLLLWVLLLWVPGSTGDGGSSRSSENLYFQGAAELTLTQSPGTL ALSPGERATLS CRAS Q S VS SNYL A WYQQKPGQ APRLLI Y GAS SRAPGIPDRF S GS GS GTDFTLTITSLQ AE D VAVYY CQ Q YFD SPLTF GGGTKVEIKGGS SRS S S S GGGGS GGGGQ V QL V Q S GAEVKKP GASVKVSCKASGYTFSSYDINWVRQATGQRPEWMGWMNPKSGDTGYAQTFQGRVTM TRSTSISTAYMELSSLRSEDTAMYYCTRGNRPIFDLFPFDLWGQGTLITVSSASTKGPSV T S GQ AGQHHHHHHGA YP YD VPD YAS GGS SRS S S S GGGGS GGGGL AE AAAKE AAAKE AA AKE AAAKE AAAKAAAGGS SRS S TS GDDDDK AKKKLPKCQKQEDCGS WDLKCNNYTK KCECRN Q V CGRGCPKERY QRDKY GCRKCLCKGCDGFKCRLGCTY GFKTDKKGCE AF C TCNTKETACV IWCTD PYKCNPESGRCEDPNEEYE YD YEGGS SRS S

1048nvw Amino Acid SEQ P ⁾ NO: 76 MKKT AI AI AV AL AGF AT V AQ AAAS S SRCT YDHWCSHGGS SRS S S S GGGGS GGGGELTL TQSPGTLALSPGERATLSCRASQSVSSNYLAWYQQKPGQAPRLLIYGASSRAPGIPDRFS GS GS GTDFTLTITSLQ AED V AVYY CQQ YFD SPLTF GGGTKVEIKGGS SRS S S S GGGGS GG GGQ V QL V Q S GAEVKKPGAS VK V S CKAS GYTF S S YDINWVRQ AT GQRPEWMGWMNPK SGDTGYAQTFQGRVTMTRSTSISTAYMELSSLRSEDTAMYYCTRGNRPIFDLFPFDLWG QGTLITVSSASTKGPSVTSGQAGQHHHHHHGAYPYDVPDYAS

Example 1 -BP230-positive sera show linear basement membrane zone immunofluorescence staining on both monkey esophagus and normal human skin sections whereas most BP sera positive only for BP180-NC16A antibodies stain only normal human skin

Sera from patients with BP230 antibodies only or both BP230 and BP180-NC16A antibodies showed linear basement membrane zone (BMZ) staining at the dermal-epidermal junction (DEJ) on both ME and NHS (Table 1). In contrast, most sera (10/12) from patients with only BP180-NC16A antibodies showed no IF signal on ME, but always on NHS. These data suggest that the BP180-NC16A is not expressed well or not expressed at all in ME. A minority of BP sera with only BP180-NC16A antibodies (2/12) did however show linear staining in ME, likely indicating presence of additional antibodies against stretches of BP 180 or BP230 not covered by the recombinant fragments used for coating of commercial ELISA plates. To study this seemingly incongruent issue in more detail we affinity-purified BP180-NC16A- and BP230- C-specific antibodies and performed additional experiments with sera depleted of both BP180- NC16A- and BP230-C-specific antibodies.

Example 2- Affinity-purified anti-BP180-NC16A polyclonal antibodies show BMZ staining on sections of normal human skin but not on monkey esophagus

Antigen-specific antibodies against BP180-NC16A and BP230-C were affinity purified from two BP patient sera. To evaluate the effectivity of affinity purification (AP), ELISA was conducted with the original BP sera (before AP), the eluates from AP, and the sample flow throughs. As illustrated in Figures 1A-1D, all APs depleted sera specifically for the antigen reactivity chosen, resulting in mono-specific eluates for downstream testing.

IIF was performed on NHS and ME substrates using BPl 80-NCl6A-/BP230-C-specific polyclonal antibodies from above APs. As already suggested by the findings from Table 1, eluates from BP230-C-APs stained both NHS and ME substrates (Figures 2A, 2B), whereas eluates from BPl80-NCl6A-APs stained the DEJ on NHS only, but not on ME (Figures 2C, 2D). Thus, for detection of pathophysiologically relevant BPl80-NCl6A-specific antibodies, NHS is a more sensitive substrate than ME. Interestingly, after double-depletion of BP180- NC16A and BP230-C specific antibodies from the same sera, linear staining at the DEJ can still be seen on both NHS and ME substrates (Figures 4A-4D), suggesting additional reactivities against epitopes outside of the BP180-NC16A and BP230-C domains that result in positive linear IIF staining. Of note, in the two sera studied here, these non-BPl80-NCl6A and non- BP230-C antibodies however did not contribute to the majority of the IIF titer.

Example 3- Monoclonal antibodies cloned from an active BP patient confirm that monkev esophagus is not bound bv anti-NCl6A reactive antibodies

ELISA testing results for four monoclonal antibodies (mAbs) obtained from antibody phage display cloning and recombinant expression are shown in Figure 3A. Immunofluorescence analysis of anti-BP230 mAbs (clones BP230-C-mAbl and BP230-C-mAb2) revealed positive staining along the BMZ on both NHS and ME (Figure 3B). IIF testing on ME was negative for anti-NCl6A mAbs BPl 80-NCl6A-mAbl and BPl80-NCl6A-mAb2, whereas on NHS linear BMZ staining was found (Figure 3C). Taken together with the results from full serum immunofluorescence testing and affinity-purified polyclonal serum antibody testing, it appears that ME is likely to be devoid of human NC16A epitopes.

The observations from Figures 2 and 3 were confirmed by performing double IIF stainings of both NHS and ME substrates with affinity-purified polyclonal anti-NCl6A-specific antibodies affinity purified from two patients (BP-l and BP-2) and the recombinant monoclonal anti-NCl6A antibody clone 1 (BPl80-NCl6A-mAbl in Figure 3A) (Figure 5).

Example 4- BP180-NC16A is genetically different in monkeys and humans

Although humans and monkeys are both primates, genetic differences exist which result in different coding amino acids (aa) in the NC16A domains of BP180. Highest homology to human NC16A is found in Pongo abelii and Gorilla gorilla gorilla, with 97% identity (Table 2A); commercially used ME usually comes from Rhesus macaques (e.g., Macaca mulatta personal communication), which has only 71% homology. Interestingly, BP230 and the non- NC16A stretches of the BP180 protein have much higher homologies between monkeys and humans, which is in line with our experimental IF findings (Figure 2B; Figure 3B; Figure 4B and 4D; Table 2B) and previous observations. Example 5- ScFv monoclonal antibodies cloned from human individuals with clinically active bullous pemphigoid react with the basement membrane zone of skin

Based on published protocols a modified antibody phage display with nested PCR was applied to cDNA isolated from peripheral blood mononuclear cells (PBMC) from two clinically active bullous pemphigoid patients and resulted in 35 monoclonal antibodies (mAbs) in the format of single chain variable fragments (scFvs) (Table 3, Table 7). Specific, custom primers were applied to obtain variable heavy chain transcripts specific for the IgG and the IgE antibody repertoires (Table 8); a nested PCR approach was applied to ensure that PCR products code specifically for the IgE or the IgG isotype. All resulting monoclonal antibodies were Sanger sequenced (for their variable heavy and light chain sequences, coding antigen-specificity) and grouped by clonality as evidenced by their heavy chain complementarity region 3 (H-CDR3) amino acid (aa) sequence; 29 unique clones were obtained applying sorting by H-CDR3 aa sequence and 80% aa similarity in between clones (mAbs from same B-cell clones are 4-5G.230 and 4-8G.230; 3-5E, 3-9E, 3-38E and 3-48E; 2-6GL and 3-29GF). Recombinantly expressed scFv proteins harbor a 6His tag and a HA tag; scFvs were isolated by their 6His tag from lysates of transformed E.coli and tested by anti-HA ELISA on BP180-NC16A and BP230 substrates; of the 29 unique clones identified, 23 bound BP180-NC16A, and 6 BP230 (Table 9, Figures 8A and 8B). Additional testing was performed by indirect immunofluorescence on normal human skin substrate for all scFvs isolated (Figures 9A-9C). For selected antibodies, western blotting on recombinant NC16A substrate and anti-Ml3 phage ELISA was performed (Table 3). All results and mAh L-/H-CDR1/2/3 aa sequences are summarized in Table 3 and Table 7.

6- Anti-BP 180-NC16A scFv antibodies are non in a model system of the human skin

Recombinant, purified and dialysed antibodies were injected into human skin organ cultures, a model system of the human skin, to access their pathogenic potential. As expected, due to absence of a cell-engaging Fc part, tested scFvs against BP180-NC16A (i.e., 3-4E, 3-16E, 3-3E, 3-14G) were not pathogenic in this assay (Table 10). Anti-BP230 scFv antibodies were not tested here, because no binding is expected to intact keratinocytes in the skin organ culture model, because of physiologic intracellular localization of this antigen. Only a mixture of scFvs (3-3E, 3-4E, 3-16E; 4:4: 1 : 1) plus a polyclonal rabbit anti-HA antibody (Sigma-Aldrich, cat. -no. H6908) which crosslinks the skin-bound scFvs by means of their HA tags resulted in

microblisters (Figure 10, asterisk), indicating that cross-linking of BP180 is required for pathogenicity, and that scFvs alone do not result in skin pathogenicity by binding to the BMZ (Figure 10, arrowheads).

Example 7- Recombinant anti-BPl80-NCl6A scFv antibodies feature displacing activity on polyclonal sera from bullous pemphigoid patients

Unexpectedly, anti-BPl 80-NCl6A mAbs (in form of scFvs) featured displacing activity on full human BP serum IgG antibodies in vitro (Figures 11A-11C). After adjusting patient sera from three BP patients (ID numbers 3419, 3389, 3391) to an OD of 0.3 to 0.5 using commercial anti- BP180-NC16A EFISA wells (EUROIMMUN, Fuebeck, Germany), differing amounts of the mAbs 3-14G (ID#l), 2-6GF (ID#29), 3-3E (ID#l6) and 4-8E. 230 (ID#28; anti-BP230) were added after a brief wash and incubated in-well. After 2 hours at 37°C, two brief washes were performed and identical, new scFv preparations were loaded again to the respective EFISA wells. Displacement of full BP serum IgG antibodies was then detected by developing with anti human IgG HRP. For serum from patient 3419, a dose-dependent, displacing activity was detected for all anti-BPl80 NC16A scFvs (ID#l, 29, 16), but not for the anti-BP230 scFv (ID#28), which served as a negative, internal control (Figure 11A). For serum from patient 3389, mAbs #1 and 16 had displacing acitivities, but not mAbs #29 and 28 (Figure 11B).

Finally, for patient’s 3391 serum, from which most of the mAbs were cloned, similar results compared to 3389 were obtained, which mAbs #1 and 16 showing displacing activity on full IgG antibodies (Figure 11C). These results are suggesting that mAbs #1 and 16 are showing displacing activity across patients (in all three sera tested), whereas mAb #29 is only displacing anti-BPl 80 NC16A IgG antibodies in the same patient’s serum (i.e., 3419), suggesting a particular antibody epitope not shared over patients. MAbs #1 and 16 may, in contrast, bind to important epitopes shared over patients, suggesting effective use in patients with BP.

Example 8- Examples of fusion proteins generated for directed therapy of diseases of skin and eve

Because most, if not all patients with BP feature complement deposits at the BMZ, resulting in linear staining in direct immunofluorescence (DIF) when stained for complement component 3 (C3), it was possible that the displacing activity of the scFvs could be combined and extended with a complement-inhibiting function for potential therapeutic use. For BP, the classical pathway of complement activation is thought to be most critical.

Fusions of the mAb#l (3-14G) with the molecularly characterized Cls-inhibiting gigastasin molecule were designed, which potentially inhibit the classical pathway of complement activation.

The first fusion compound designed was l005sqv, in which the gigastasin-domain was N terminal, followed by a linker and the scFv-domain 3-14G (Figure 12A). As a signal sequence for eukaryotic expression, the murine Ig kappa chain V-III region signal sequence was chosen because of its confirmed and robust use in protein expression. Despite good expression yields and reactivity in the anti-HA-EFISA on BP180-NC16 substrate (Figure 12B and 12C), this compound did not bind to the BMZ of human skin cryosections, as evidenced by indirect immunofluorescence (Figure 12D). Despite this finding, it did inhibit complement fixation by BP antibodies, as evidenced by the routine in vitro complement-fixation test, likely because of the presence of the unbound compound in the reaction supernatant (Figure 12E).

To ultimately obtain targeted binding of a Cls-inhibiting fusion compound to the BMZ of human skin, the compound was redesigned as outlined in Figure 13; with an N-terminal position of the BMZ-binding anti-BPl80-NCl6A domain 3-14G and a C-terminal position of the gigastasin domain (separated by a helical linker to obtain physical separation in between the two domains; fusion name l0l2qxm; Figure 13A). Good expression efficacy was observed in eukaryotic cells (Figure 13B), as well as binding to BP180-NC16A substrate by anti-HA ELISA (Figure 13C). Binding to the BMZ of human skin (Figure 13D) was also evaluated by indirect immunofluorescence. As expected, the fusion compound l0l2qxm resulted in inhibition of C3 fixation at the BMZ in the CFT, whereas the 3-14G scFv alone did not result in inhibition of C3 fixation (Figure 13E); these outcomes were then independently confirmed by assessment of the anaphylatoxin C5a (a marker of complement cascade activation further downstream of C3) in the reaction supernatants, with l0l2qxm leading to a significant decrease in C5a liberation in vitro when compared to the 3-14G scFv alone (mean optical densities, ODs, of 1.371±0.2193 vs. 0.4209±0.0452; p=0.0444 in the unpaired t test with Welch’s correction; Figure 13F).

To assess the potential effect of inhibition of the alternative pathway of complement activation, another compound, l048nvw, was designed that combines the factor H-binding 5C6 peptide ASSSRCTYDHWCSH (Wu _. et al. _. , _. 201 _. 1 _. ) with the BMZ-targeting 3-14G scFv domain in one fusion protein (Figure 14A): Because of the small size of the 5C6 domain, the difference in size is small when compared to the 3-14G scFv molecule alone by SDS-PAGE (Figure 14B; lanes 5 and 6); this relatively small size was also the reason that expression was done in E.coli, with the ompA signal peptide incorporated. In anti-HA ELISA, the fusion compound shows high and dose-dependent reactivity on BP180-NC16A substrate (Figure 14C). As evaluated by indirect immunofluorescence, the fusion l048nvm binds to the BMZ (anti-HA staining in Figure 14D) and binds factor H (FH) from the pooled plasma in the supernatant (anti-FH staining in Figure 14D; Cat. No. A312, Quidel). However, the complement-inhibiting effect is small, if present at all, when evaluated by anti-C3 staining in the CFT (Figure 14E), suggesting that the classical complement activation pathway is contributing most to the observed complement fixation at the BMZ. Table 1-Twenty-nine BP patients from the Shanghai Department of Dermatology at the Rui Jin Hospital, Shanghai Jiao Tong University School of medicine, Shanghai, China, were assayed by routine ELISA testing and by IIF on monkey esophagus (ME) and normal human skin (NHS). Titers tested by IIF testing on ME and NHS were 1 : 100 for all sera and, additionally, 1 : 10 for cases with negative IIF readings at 1 : 100.

Table 2A-Genetic comparison of human BP180, its NC16A domain, and BP230, over different species.

Human BP230: NCBI Reference Sequence NP_001714.1

Table 2B-Titers on NHS and ME before and after double depletion. Most antibodies contributing to the IF titer are removed by depletion on BP 180-NC16A and BP230-C antigen. The resultant non-BPl 80-NC16A and/or non-BP230-C antibodies contribute to a minor fraction of the IF on NHS only. By comparing titers before and after double depletion, their share appear to be larger on ME substrate, likely because BP180-NC16A antibodies are not contributing to an

IF signal here in the first place, i.e., before double depletion.

Table 3

Table 4-results shown in Figure 11 A.

all scFv dilutions adjusted to 1 ug/uL stock cone. Table 5-results shown in Figure 11B.

all scFv dilutions adjusted to 1 ug/uL stock cone.

Table 6-Results shown in Figure 11C.

all scFv dilutions adjusted to 1 ug/uL stock cone.

WO 2020/072937 PCT/US2019/054751

Table 7-CDRs of scFvs shown in Table 3

Table 8- Variable heavy chain PCR primers

Table 9- Results shown in Figure 8.

Table 10-Human skin organ culture results

Other Embodiments

The recitation of a listing of elements in any definition of a variable herein includes definitions of that variable as any single element or combination (or subcombination) of listed elements. The recitation of an embodiment herein includes that embodiment as any single embodiment or in combination with any other embodiment or portions thereof.

The disclosures of each and every patent, patent application, and publication cited herein are hereby incorporated herein by reference in their entirety. While this invention has been disclosed with reference to specific embodiments, it is apparent that other embodiments and variations of this in vention may be devised by others skilled in the art without departing from the true spirit and scope of the invention. The appended claims are intended to be construed to include all such embodiments and equivalent variations.