Field of the invention

The present invention relates to an antibody composition, preferably an IgY antibody composition, and the use of such a composition for diagnosis and prevention and/or treatment of disease caused by an infection of a coronavirus or other RNA viruses.

Background of the invention

Coronaviruses are a group of related RNA viruses that cause diseases in mammals and birds. In humans, the infection occurs by virus binding to cell receptors on epithelial cells which enables the virus particles to enter the cells. The principal mode by which humans are infected with coronavirus is through exposure to respiratory fluids carrying infectious virus. Exposure occurs in three principal ways: (1) inhalation of very fine respiratory droplets and aerosol particles, (2) deposition of respiratory droplets and particles on exposed mucous membranes in the mouth, nose, or eye by direct splashes and sprays, and (3) touching mucous membranes with hands that have been soiled either directly by viruscontaining respiratory fluids or indirectly by touching surfaces with virus on them. Human coronaviruses are capable of causing illnesses ranging from the common cold to more severe diseases such as Middle East respiratory syndrome (MERS, fatality rate -34%). SARS-CoV-2 is the seventh known coronavirus to cause respiratory diseases in humans after 229E, NL63, OC43, HKU1, MERS- CoV, and the original SARS-CoV.

In December 2019, a pneumonia outbreak was reported in Wuhan, China. On 31 December 2019, the outbreak was traced to a novel strain of coronavirus, which was given the interim name 2019-nCoV by the World Health Organization (WHO), later renamed SARS-CoV-2 by the International Committee on Taxonomy of Viruses. Angiotensin converting enzyme 2 (ACE2), a host epithelial cell receptor is found to be the main entry point for SARS-CoV-2. Neuropilin-1 (NRP-1), another host cell receptor, further potentiates SARS-CoV-2 infectivity. Coronavirus disease 2019 (COVID-19) is defined by WHO as illness caused by SARS-CoV-2. People with COVID-19 have had a wide range of symptoms reported - ranging from mild symptoms to severe illness. Of those people who develop noticeable symptoms 81% develop mild to moderate symptoms (up to mild pneumonia), while 14% develop severe symptoms (dyspnea, hypoxia, or more than 50% lung involvement on imaging), and 5% suffer critical symptoms (respiratory failure, shock, or multiorgan dysfunction). Older people are at a higher risk of developing severe symptoms. Some people continue to experience a range of effects (long COVID) for months after recovery, and damage to organs has been observed.

Since then, vaccines and treatments for COVID-19 have been developed, but there is still a need to explore new ways to treat and prevent human coronavirus infections by developing new and improved antibodies, drugs, and vaccines.

Objects of the invention

An object of the present invention is to develop antibody compositions that are specific to corona viruses or other respiratory viruses, in particular the SARS-CoV-2 virus, in addition to surface receptors angiotensin-converting enzyme 2 (ACE-2) and/or neuropilin 1 (NRP-l) expressed on human cells, as well as nucleic acids encoding said antibodies.

An object of the present invention is to develop polyclonal antibodies that are specific to corona viruses or other respiratory viruses, and which also is specific to the SARS-CoV-2 virus virus, in addition to surface receptors angiotensin-converting enzyme 2 (ACE-2) and neuropilin 1 (NRP-l) expressed on human cells.

A further object of the present invention is to administer the antigens to the antibody producing animals so that the animals are immunized.

A further object of the present invention relates to the use of antibodies for use in prevention and/or treatment of an infection of a coronavirus in humans.

A further object of the present invention relates to the use of the antibody compositions for a diagnostic test.

A further object of the present invention relates to the use of polyclonal IgY antibodies for a diagnostic test.

These and other objects of the invention will be described in further detail hereinafter. Summary of the invention

In one aspect, the present invention provides a composition comprising one or more antibodies and/or antigen binding fragments, and/or one or more nucleic acids encoding any one or more of said antibodies and/or antigen binding fragments, selected from the group consisting of:

(a) one or more antibodies and/or antigen binding fragments with specificity to an ACE-2 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments;

(b) one or more antibodies and/or antigen binding fragments with specificity to an NRP-1 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments;

(c) one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 Spike 1 RBD antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments; and

(d) one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 SI antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments.

In another aspect, the present invention relates to an IgY antibody composition, wherein the IgY antibody composition is obtained by immunization of an egg-laying poultry with a single or a combination of any of the following antigens: Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-1) antigen, SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, and SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

In another aspect, the present invention relates to an IgY antibody composition for use in prevention and/or treatment of an infection of a coronavirus in an animal, wherein the IgY antibody composition is obtained by immunization of an egg-laying poultry with a single or a combination of any of the following antigens Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-1) antigen, SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, and SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

In another aspect, the present invention relates to an IgY antibody composition, wherein the IgY antibody composition is obtained by immunization of an egg-laying poultry with a single or a combination of any of the following antigens: Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-1) antigen, SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, and SARS-CoV-2 Spike Glycoprotein 1 (S 1) antigen, and at least one medical compatible adjuvant. It will be appreciated that in a passive immunization therapy in a patient, inclusion of an adjuvant might not be useful or appropriate. However, for an active immunization therapy in a patient and/or in a method of producing IgY antibodies in a layer then an adjuvant might be a useful addition.

In another aspect, the invention relates to an egg yolk from an egg from an egg-laying poultry for use in prevention and/or treatment of an infection of a coronavirus in an animal, wherein the egg yolk comprises IgY antibodies obtained by immunization of the egg-laying poultry with a single or a combination of any of the following antigens: Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-1) antigen, SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, and SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

In another aspect, the present invention relates to a method of in vitro diagnosis, wherein an IgY antibody composition, which is obtained by immunization of an egg-laying poultry with a single or a combination of any of the following antigens: Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-1) antigen, SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, and SARS-CoV-2 Spike Glycoprotein 1 (SI), is contacted with a biological sample from an animal, and wherein a neutralization or complexation of the IgY antibody composition with a virus in the sample indicates that the sample contains corona virus or other RNA viruses.

These and other aspects of the invention will be described in further detail hereinafter.

Brief description of the drawings

Figures 1 to 15 show results obtained by evaluating the present invention, as further described in the examples.

Detailed description of the invention

The aim of the present invention is to provide antibodies that are specific to corona virus or other viruses, and/or antibodies that are specific to surface receptors ACE-2 (herein also referred to as ACE2) and/or NRP-1 (herein also referred to as NRP) on animal cells, preferably human cells. Preferably, the antibodies will reduce or prevent virus infection of the host animal, by blocking binding sites on the virus or the host cell surface that the virus is dependent on for entry into host cells. Preferably, the antibodies are IgY antibodies, more preferably polyclonal IgY antibodies.

Antibody combination compositions

In a first aspect of the further combination composition embodiments, the present invention provides a composition comprising, consisting essentially of, or consisting of one or more antibodies and/or antigen binding fragments, and/or one or more nucleic acids encoding any one and/or more of said antibodies and/or antigen binding fragments, selected from the group consisting of:

(a) one or more antibodies and/or antigen binding fragments with specificity to an ACE-2 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments;

(b) one or more antibodies and/or antigen binding fragments with specificity to an NRP-1 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments;

(c) one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 Spike 1 RBD antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments; and

(d) one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 SI antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments.

In the context of the present invention, by one or more antibodies and/or antigen binding fragments or one or more nucleic acids encoding any one or more of said antibodies and/or antigen binding fragments, we include at least one, two, three or four different forms of antibodies and/or antibody binding fragments as specified in parts (a), (b), (c) and/or (d) of the preceding paragraph, and/or at least one, two, three or four different forms of nucleic acids encoding said antibodies and/or antigen binding fragments as specified in parts (a), (b), (c) and/or (d) of the preceding paragraph. A form of antibodies and/or antibody binding fragments against a particular antigen can include, for example a population of antibody molecules, and/or a population of antibody binding fragments thereof, which population can be either homogeneous or heterogeneous, and which may for example comprise, consist essentially of, or consist of a population of antibody molecules, and/or a population of antibody binding fragments thereof, having specificity for the same antigen, and/or for the same epitope. Exemplary forms of such antibody populations include polyclonal and monoclonal preparations, including polyclonal preparations which can be enriched for antibodies (or fragments thereof) with binding specificity to a defined antigen and/or a defined epitope. Polyclonal antibody preparations are considered heterogenous in that multiple antibodies, having different CDR sequences, are typically present and able to bind to the defined antigen and/or a defined epitope. Said populations, if heterogenous, may or may not additionally include other forms of antibody, and/or antibody binding fragments thereof, with different binding specificities (i.e. which do not bind specifically to the defined antigen and/or the defined epitope), provided that said population contains (e.g. comprises or consists essentially of) the antibodies and/or antibody binding fragments against the defined antigen and/or the defined epitope, typically in an effective amount. In the following passages, references to an “antibody” can optionally also include reference to a “form of antibodies” and references to fragments thereof can also optionally include reference to a “form of fragments thereof’.

In a particular embodiment, the composition comprises one or more antibodies and/or antigen binding fragments with specificity to an ACE-2 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments.

An example of an ACE-2 antigen may have the amino acid sequence of SEQ ID NO: 1, below - Accession # Q9BYF1-2. A particular example of ACE-2 antigen is available from The Native Antigen Company, which includes amino acids 18-615 and/or AcroBiosystems which includes amino acids 18- 740.

MSSSSWLLLSLVAVTAAQSTIEEQAKTFLDKFNHEAEDLFYQSSLASWNYNTNITEE NVQNM NNAGDKWSAFLKEQSTLAQMYPLQEIQNLTVKLQLQALQQNGSSVLSEDKSKRLNTILNT M STIYSTGKVCNPDNPQECLLLEPGLNEIMANSLDYNERLWAWESWRSEVGKQLRPLYEEY V VLKNEMARANHYEDYGDYWRGDYEVNGVDGYDYSRGQLIEDVEHTFEEIKPLYEHLHAYV RAKLMNAYPSYISPIGCLPAHLLGDMWGRFWTNLYSLTVPFGQKPNIDVTDAMVDQAWDA QRIFKEAEKFFVSVGLPNMTQGFWENSMLTDPGNVQKAVCHPTAWDLGKGDFRILMCTKV T MDDFLTAHHEMGHIQYDMAYAAQPFLLRNGANEGFHEAVGEIMSLSAATPKHLKSIGLLS P DFQEDNETEINFLLKQALTIVGTLPFTYMLEKWRWMVFKGEIPKDQWMKKWWEMKREIVG VVEPVPHDETYCDPASLFHVSNDYSFIRYYTRTLYQFQFQEALCQAAKHEGPLHKCDISN STE AGQKLFNMLRLGKSEPWTLALENVVGAKNMNVRPLLNYFEPLFTWLKDQNKNSFVGWSTD WSPYADQSIKVRISLKSALGDKAYEWNDNEMYLFRSSVAYAMRQYFLKVKNQMILFGEED V RVANLKPRISFNFFVTAPKNVSDIIPRTEVEKAIRMSRSRINDAFRLNDNSLEFLGIQPT LGPPN QPPVSIWLIVFGVVMGVIVVGIVILIFTGIRDRKKKNKARSGENPYASIDISKGENNPGF QNTD DVQTSF (SEQ ID NO: 1)

In a particular embodiment, the ACE-2 antigen comprises SEQ ID NO: 1, or an immunogenic fragment thereof and/or an immunogenic variant thereof.

By “immunogenic fragment”, we include a fragment of the antigen (such as the ACE-2, NRP-1, RBD, and/or S 1 antigens) that is capable of inducing an immune response, wherein that immune response would be comparable or the same as would be induced to the antigen. The fragment may lack one or more amino acids at the N-terminus, C-terminus and/or in the center of the antigen.

By “immunogenic variant”, we include a variant of the antigen (for example, a version of the antigen with one or more mutations) that is capable of inducing an immune response, and wherein that immune response would be comparable or the same as would be induced to the antigen.

In a particular embodiment, the immunogenic fragment comprises 5 or more amino acids of the antigen; for example: about 10 or more; about 20 or more; about 30 or more; about 40 or more; about 50 or more; about 60 or more; about 70 or more; about 80 or more; about 90 or more; about 100 or more; about 110 or more; about 120 or more; about 130 or more; about 140 or more; about 150 or more; about 160 or more; about 170 or more; about 180 or more; about 190 or more; about 200 or more; about 250 or more; about 300 or more; about 350 or more; about 400 or more; about 450 or more; about 500 or more amino acids of the antigen. In a particular embodiment, the immunogenic fragment comprises about 5 to about 50 amino acids of the antigen; about 5 to about 500 amino acids of the antigen; about 100 to about 500 amino acids of the antigen.

In particular embodiments, the immunogenic variant and/or immunogenic fragment is more than about 50%, about 60%, about 70%, about 75%, such as more than about 80%, about 85%, more than about 90%, or even more preferably more than about 95% or about 99% identical to the antigen (such as the ACE-2, NRP-1, RBD, and/or SI antigens).

The percent identity between two amino acid sequences is determined as follows. First, an amino acid sequence is compared to, for example, SEQ ID NO: 1 using the BLAST 2 Sequences (B12seq) program from the stand-alone version of BLASTZ containing BLASTN version 2.0.14 and BLASTP version 2.0.14. This stand-alone version of BLASTZ can be obtained from the U.S. government’s National Center for Biotechnology Information web site at ncbi.nlm.nih.gov. Instructions explaining how to use the B12seq program can be found in the readme fde accompanying BLASTZ. B12seq performs a comparison between two amino acid sequences using the BLASTP algorithm. To compare two amino acid sequences, the options of B12seq are set as follows: -i is set to a file containing the first amino acid sequence to be compared (e.g., C:\seql.txt); -j is set to a file containing the second amino acid sequence to be compared (e.g., C:\seq2.txt); -p is set to blastp; -o is set to any desired file name (e.g., C:\output.txt); and all other options are left at their default setting. For example, the following command can be used to generate an output file containing a comparison between two amino acid sequences: C:\B12seq -i c:\seql.txt -j c:\seq2.txt -p blastp -o c:\output.txt. If the two compared sequences share homology, then the designated output file will present those regions of homology as aligned sequences. If the two compared sequences do not share homology, then the designated output file will not present aligned sequences. Once aligned, the number of matches is determined by counting the number of positions where an identical nucleotide or amino acid residue is presented in both sequences.

The percent identity is determined by dividing the number of matches by the length of the sequence set forth in an identified sequence followed by multiplying the resulting value by 100.

In one embodiment, the ACE-2 immunogenic fragment is amino acids 18-615 of SEQ ID NO: 1 and/or amino acids 18-740 of SEQ ID NO: 1.

In a particular embodiment, the composition comprises one or more antibodies and/or antigen binding fragments with specificity to an NRP-1 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments.

An example of an NRP-1 antigen may have the amino acid sequence of SEQ ID NO: 2, below - Accession # AAH07533. A particular example ofNRP-1 antigen is available from 2bScientific, which is human NRP1, with an Fc Tag (NR1-H5252) and is expressed from human 293 cells (HEK293) and contains AA Phe 22 - Lys 644.

MERGLPLLCAVLALVLAPAGAFRNDKCGDTIKIESPGYLTSPGYPHSYHPSEKCEWL IQAPDP YQRIMINFNPHFDLEDRDCKYDYVEVFDGENENGHFRGKFCGKIAPPPVVSSGPFLFIKF VSD YETHGAGFSIRYEIFKRGPECSQNYTTPSGVIKSPGFPEKYPNSLECTYIVFAPKMSEII LEFESF DLEPDSNPPGGMFCRYDRLEIWDGFPDVGPHIGRYCGQKTPGRIRSSSGILSMVFYTDSA IAK EGFSANYSVLQSSVSEDFKCMEALGMESGEIHSDQITASSQYSTNWSAERSRLNYPENGW TP GEDSYREWIQVDLGLLRFVTAVGTQGAISKETKKKYYVKTYKIDVSSNGEDWITIKEGNK PV LFQGNTNPTDVVVAVFPKPLITRFVRIKPATWETGISMRFEVYGCKITDYPCSGMLGMVS GLI SD SQITS SNQGDRNWMPENIRLVTSRSGWALPPAPHSYINEWLQIDLGEEKIVRGIIIQGGKHR ENKVFMRKFKIGYSNNGSDWKMIMDDSKRKAKSFEGNNNYDTPELRTFPALSTRFIRIYP ER ATHGGLGLRMELLGCEVEAPTAGPTTPNGNLVDECDDDQANCHSGTGDDFQLTGGTTVLA T EKPTVIDSTIQSGIK (SEQ ID NO: 2)

A further example of an NRP-1 antigen may have the amino acid sequence of SEQ ID NO: 3, below - Accession 014786 (NRPI HUMAN):

MERGLPLLCAVLALVLAPAGAFRNDKCGDTIKIESPGYLTSPGYPHSYHPSEKCEWL IQAPDP YQRIMINFNPHFDLEDRDCKYDYVEVFDGENENGHFRGKFCGKIAPPPVVSSGPFLFIKF VSD YETHGAGFSIRYEIFKRGPECSQNYTTPSGVIKSPGFPEKYPNSLECTYIVFVPKMSEII LEFESF DLEPDSNPPGGMFCRYDRLEIWDGFPDVGPHIGRYCGQKTPGRIRSSSGILSMVFYTDSA IAK EGFSANYSVLQSSVSEDFKCMEALGMESGEIHSDQITASSQYSTNWSAERSRLNYPENGW TP GEDSYREWIQVDLGLLRFVTAVGTQGAISKETKKKYYVKTYKIDVSSNGEDWITIKEGNK PV LFQGNTNPTDVVVAVFPKPLITRFVRIKPATWETGISMRFEVYGCKITDYPCSGMLGMVS GLI SD SQITS SNQGDRNWMPENIRLVTSRSGWALPPAPHSYINEWLQIDLGEEKIVRGIIIQGGKHR ENKVFMRKFKIGYSNNGSDWKMIMDDSKRKAKSFEGNNNYDTPELRTFPALSTRFIRIYP ER ATHGGLGLRMELLGCEVEAPTAGPTTPNGNLVDECDDDQANCHSGTGDDFQLTGGTTVLA T EKPTVIDSTIQSEFPTYGFNCEFGWGSHKTFCHWEHDNHVQLKWSVLTSKTGPIQDHTGD GN

FIYSQADENQKGKVARLVSPVVYSQNSAHCMTFWYHMSGSHVGTLRVKLRYQKPEEY DQL VWMAIGHQGDHWKEGRVLLHKSLKLYQVIFEGEIGKGNLGGIAVDDISINNHISQEDCAK PA DLDKKNPEIKIDETGSTPGYEGEGEGDKNISRKPGNVLKTLDPILITIIAMSALGVLLGA VCGV VLYCACWHNGMSERNLSALENYNFELVDGVKLKKDKLNTQSTYSEA (SEQ ID NO: 3)

In a particular embodiment, the NRP-1 antigen comprises SEQ ID NO: 2 or SEQ ID NO: 3 (preferably SEQ ID NO: 3), or an immunogenic fragment and/or variant thereof.

In a particular embodiment, the composition comprises one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 Spike 1 RBD antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments. An example of a SARS-CoV-2 RBD antigen may have the amino acid sequence below - NCBI Accession Number: 6XDG E. A particular example of RBD antigen is SEQ ID NO: 4 available from The Native Antigen Company, which is a recombinant SARS-CoV-2 Spike SI RBD (aa 319-541), manufactured in HEK293 cells with His-tag.

RVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFS TFKCYG VSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNL DS KVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGV GY QPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFEQKLISEEDLGGEQKLISEEDLHH HH HH (SEQ ID NO: 4)

A further example of an RBD antigen may have the amino acid sequence of SEQ ID NO: 5, below - Accession P0DT2 (amino acids 319-541 of SI), which is wild type Wuhan RBD sequence:

RVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFS TFKCYG VSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNL DS KVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGV GY QPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNF (SEQ ID NO: 5)

However, it will be appreciated that the RBD antigen from various strains (also referred to as variants) of SARS-CoV-2 could also be used as part of the invention; for example, the Epsilon, Eta, Iota, Kappa, Alpha, Beta, Delta, Gamma and/or Lambda variants, preferably the Delta variant. These above known variants have the mutations shown below in Table A. SARS-CoV-2 variants are continuously arising, including those that may have new mutations in the RBD domain. It will be appreciated by the skilled person that any future SARS-CoV-2 variants with mutated RBD domains are included in the present invention, such that antibodies and/or antigen-binding fragments with specificity to such mutated RBD antigens can be included herein, and such mutated RBD antigens can be used to generate IgY antibodies through the immunization of layers as discussed herein.

Table A, the RBD mutations, when compared to the Wuhan RBD sequence (x* = occurs in some sequences but not all):

In a particular embodiment, the RBD antigen comprises SEQ ID NO: 4 and/or SEQ ID NO: 5 (preferably SEQ ID NO: 5), or an immunogenic fragment and/or variant thereof.

In a further embodiment, the RBD antigen variant comprises the amino sequence of SEQ ID NO: 4 and/or SEQ ID NO: 5 (preferably SEQ ID NO: 5), in which one or more of the mutations of Table A is applied.

In a particular embodiment, the composition comprises one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 SI antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments.

An example of a SARS-CoV-2 S 1 antigen may have the amino acid sequence of SEQ ID NO: 6, below - this is from Wuhan-Hu-1 strain, NCBI Accession Number: YP 009724390.1. A further example of a SI antigen is residues 1-674 of SEQ ID NO: 6 that has the amino acid sequence of SEQ ID NO: 7, below. However, it will be appreciated that the SI antigen from other strains of SARS-CoV-2 could also be used as part of the invention. A particular example of SI is residues 1-674 of SEQ ID NO: 6 (i.e. SEQ ID NO: 7), C-terminally tagged with sheep Fc as available from The Native Antigen Company, which is a recombinant protein manufactured in HEK293 cells and purified from culture supernatant by Protein G chromatography. MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFS NVT WFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNA TNV VIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNF K NLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRS YLTPG DSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGI YQ TSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASF STFK CYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNS NN LDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPT NGV GYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQ QF GRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIH ADQ LTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRARSVA SQSI IAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNL LLQY GSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRS FIEDL LFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGT ITS GWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASA LG KLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDKVEAEVQIDRLITGRLQSLQT YVT QQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVP A QEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVI GIV NNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNL NES LIDLQELGKYEQYIKWPWYIWLGFIAGLIAIVMVTIMLCCMTSCCSCLKGCCSCGSCCKF DED DSEPVLKGVKLHYT (SEQ ID NO: 6)

MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLP FFSNVT WFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNA TNV VIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNF K NLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRS YLTPG DSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGI YQ TSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASF STFK CYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNS NN LDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPT NGV GYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQ QF GRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIH ADQ LTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASY (SEQ ID NO: 7) A further example of an SI antigen may have the amino acid sequence of SEQ ID NO: 8, below - Accession UniProtKB - A0A6H1PJZ3 (A0A6H1PJZ3 SARS2) (the last four amino acids RRAR is the Furin Cleavage site which is where spike is cleaved into SI and S2):

MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLP FFSNVT WFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNA TNV VIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNF K NLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRS YLTPG DSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGI YQ TSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASF STFK CYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNS NN LDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPT NGV GYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQ QF GRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQGVNCTEVPVAIH ADQ LTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRAR (SEQ ID NO: 8)

In a particular embodiment, the SI antigen comprises SEQ ID NO: 6 and/or SEQ ID NO: 7 and/or SEQ ID NO: 8 (preferably SEQ ID NO: 8), or an immunogenic fragment and/or variant thereof.

In a further embodiment, the SEQ ID NO: 8 does not comprise the C-terminal RRAR amino acids.

Without wanting to be bound by theory, based on the teaching in the Examples it is considered that for polyclonal antibodies with specificity to SI the most therapeutically functional antibodies within that population are likely to those that bind to the RBD domain within SI, whilst other antibodies in the polyclonal antibody population that bind other parts of S 1 may be less functional from the therapeutic perspective.

As described above in respect of the RBD antigen, S 1 antigen from various strains (also referred to as variants) of SARS-CoV-2 could also be used as part of the invention; for example, the Epsilon, Eta, Iota, Kappa, Alpha, Beta, Delta, Gamma and/or Lambda variants, preferably the Delta variant. SARS- CoV-2 variants are continuously arising, including those that may have new mutations in S 1. It will be appreciated by the skilled person that any future SARS-CoV-2 variants with mutated SI are included in the present invention, such that antibodies and/or antigen-binding fragments with specificity to such mutated SI antigens can be included herein, and such mutated SI antigens can be used to generate IgY antibodies through the immunization of layers as discussed herein.

In one embodiment, the antigen (such as the ACE-2, NRP-1, SI and/or RBD antigen) is recombinant.

In one embodiment, the composition is for passive immunization.

In one embodiment, the composition comprises, consists essentially of, or consists of two or more antibodies and/or antigen binding fragments, and/or two or more nucleic acids encoding any two or more of said antibodies and/or antigen binding fragments, selected from the group consisting of:

(a) one or more antibodies and/or antigen binding fragments with specificity to an ACE-2 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments;

(b) one or more antibodies and/or antigen binding fragments with specificity to an NRP-1 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments;

(c) one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 Spike 1 RBD antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments; and

(d) one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 SI antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments, optionally wherein the composition comprises a combination of the two and/or more antibodies antigen binding fragments, and/or two or more nucleic acids encoding any two and/or more of said antibodies antigen binding fragments, selected from the list consisting of: (a) and (b); (a) and (c); (a) and (d); (b) and (c); (b) and (d); (c) and (d); (a), (b) and (c); (a), (b) and (d); (a), (c) and (d); (b), (c) and (d); and (a), (b), (c) and (d), preferably selected from the list consisting of: (a) and (c); (a) and (d); (a), (b) and (c), and (a), (b) and (d); and (a), (c) and (d), most preferably selected from: (a) and (c); (a), (b) and (c), and (a), (b) and (d), and even more preferably (a), (b) and (c) or (a) and (c).

In a preferred embodiment, the composition comprises the following antibodies and/or antigen binding fragments, and/or nucleic acids encoding said antibodies and/or antigen binding fragments:

- one or more antibodies and/or antibody fragments thereof with specificity to an ACE-2 antigen; and - one or more antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike 1 RBD antigen.

In a more preferred embodiment, the composition comprises the following antibodies and/ or antigen binding fragments, and/or nucleic acids encoding said antibodies and/or antigen binding fragments:

- one or more antibodies and/or antibody fragments thereof with specificity to an ACE-2 antigen;

- one or more antibodies and/or antibody fragments thereof with specificity to an NRP-1 antigen; and

- one or more antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike 1 RBD antigen.

In a preferred embodiment, the composition comprises antibodies and/or antigen binding fragments.

By “an antibody and/or an antigen-binding fragment thereof’ we include substantially intact antibody molecules, as well as chimeric antibodies, humanised antibodies, isolated human antibodies, single chain antibodies, bispecific antibodies, antibody heavy chains, antibody light chains, homodimers and heterodimers of antibody heavy and/or light chains, and antigen-binding fragments and derivatives of the same. Suitable antigen-binding fragments and derivatives include Fv fragments (e.g. single chain Fv and disulphide-bonded Fv), Fab-like fragments (e.g. Fab fragments, Fab’ fragments and F(ab)2 fragments), single variable domains (e.g. VH and VL domains) and single domain antibodies (dAbs, including single and dual formats [i.e. dAb-linker-dAb], and nanobodies). The potential advantages of using antibody fragments, rather than whole antibodies, are several-fold. The smaller size of the fragments may lead to improved pharmacological properties, such as better penetration of solid tissue. Moreover, antigen-binding fragments such as Fab, Fv, ScFv and dAb antibody fragments can be expressed in and secreted from E. coli, thus allowing the facile production of large amounts of the said fragments.

In one embodiment, the antigen-binding fragment is selected from the group consisting of: Fv fragments (such as a single chain Fv fragment, or a disulphide-bonded Fv fragment), Fab-like fragments (such as a Fab fragment; a Fab’ fragment or a F(ab)2 fragment) and single domain antibodies.

The phrase “an antibody or an antigen-binding fragment thereof’ is also intended to encompass antibody mimics (for example, non-antibody scaffold structures that have a high degree of stability yet allow variability to be introduced at certain positions). Those skilled in the art of biochemistry will be familiar with many such molecules, as discussed in Gebauer & Skerra, 2009 (the disclosures of which are incorporated herein by reference). Exemplary antibody mimics include: affibodies (also called Trinectins; Nygren, 2008, FEBS J, 275, 2668-2676); CTLDs (also called Tetranectins; Innovations Pharmac. Technol. (2006), 27-30); adnectins (also called monobodies; Meth. Mol. Biol., 352 (2007), 95-109); anticalins (Drug Discovery Today (2005), 10, 23-33); DARPins (ankyrins; Nat. Biotechnol. (2004), 22, 575-582); avimers (Nat. Biotechnol. (2005), 23, 1556-1561); microbodies (FEBS J, (2007), 274, 86-95); peptide aptamers (Expert. Opin. Biol. Ther. (2005), 5, 783-797); Kunitz domains (J. Pharmacol. Exp. Ther. (2006) 318, 803-809); affilins (Trends. Biotechnol. (2005), 23, 514-522); affimers (Avacta Life Sciences, Wetherby, UK).

Also included within the scope of the invention are chimeric T cell receptors (also known as chimeric immunoreceptors, and chimeric antigen receptors or CARs) (see Pule et al., 2003, the disclosures of which are incorporated herein by reference). These are engineered receptors, which graft an arbitrary specificity onto an immune effector cell. Typically, CARs are used to graft the specificity of a monoclonal antibody onto a T cell; with transfer of their coding sequence facilitated by retroviral vectors. The most common form of such molecules is fusions comprising a single-chain variable fragment (scFv) derived from a monoclonal antibody fused to CD3-zeta transmembrane and endodomain. When T cells express this fusion molecule, they recognize and kill target cells that express the transferred monoclonal antibody specificity.

Persons skilled in the art will further appreciate that the invention also encompasses modified versions of antibodies and antigen-binding fragments thereof, whether existing now or in the future, e.g. modified by the covalent attachment of polyethylene glycol or another suitable polymer.

Methods of generating antibodies and antibody fragments are well known in the art. For example, antibodies may be generated via any one of several methods which employ induction of in vivo production of antibody molecules, screening of immunoglobulin libraries (Orlandi. et al, 1989; Winter et al., 1991, the disclosures of which are incorporated herein by reference) or generation of monoclonal antibody molecules by cell lines in culture. These include, but are not limited to, the hybridoma technique, the human B cell hybridoma technique, and the Epstein-Barr virus (EBV)-hybridoma technique (Kohler et al., 1975,Kozbor et al., 1985; Cote et al., 1983; Cole et al., 1984., the disclosures of which are incorporated herein by reference).

Suitable methods for the production of monoclonal antibodies are also disclosed in “Monoclonal Antibodies: A manual of techniques”, H Zola (CRC Press, 1988, the disclosures of which are incorporated herein by reference) and in “Monoclonal Hybridoma Antibodies: Techniques and Applications”, J G R Hurrell (CRC Press, 1982, the disclosures of which are incorporated herein by reference).

Likewise, antibody fragments can be obtained using methods well known in the art (see, for example, Harlow & Lane, 1988, “Antibodies: A Laboratory Manual”, Cold Spring Harbor Laboratory, New York, the disclosures of which are incorporated herein by reference). For example, antibody fragments according to the present invention can be prepared by proteolytic hydrolysis of the antibody or by expression in E. coli or mammalian cells (e.g. Chinese hamster ovary cell culture or other protein expression systems) of DNA encoding the fragment. Alternatively, antibody fragments can be obtained by pepsin or papain digestion of whole antibodies by conventional methods.

It will be appreciated by persons skilled in the art that for human therapy or diagnostics, human or humanised antibodies are preferably used. Humanised forms of non-human (e.g. murine) antibodies are genetically engineered chimaeric antibodies or antibody fragments having preferably minimal- portions derived from non-human antibodies. Humanised antibodies include antibodies in which complementary determining regions of a human antibody (recipient antibody) are replaced by residues from a complementary determining region of a non-human species (donor antibody) such as mouse, rat or rabbit having the desired functionality. In some instances, Fv framework residues of the human antibody are replaced by corresponding non-human residues. Humanised antibodies may also comprise residues which are found neither in the recipient antibody nor in the imported complementarity determining region or framework sequences. In general, the humanised antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the complementarity determining regions correspond to those of a non-human antibody and all, or substantially all, of the framework regions correspond to those of a relevant human consensus sequence. Humanised antibodies optimally 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, Riechmann et al., 1988, Presta, 1992, the disclosures of which are incorporated herein by reference).

Methods for humanising non-human antibodies are well known in the art. Generally, the humanised antibody has one or more amino acid residues introduced into it from a source which is non-human. These non-human amino acid residues, often referred to as imported residues, are typically taken from an imported variable domain. Humanisation can be essentially performed as described (see, for example, Jones et al., 1986, Reichmann et al., 1988, Verhoeyen et al., 1988, US 4,816,567, the disclosures of which are incorporated herein by reference) by substituting human complementarity determining regions with corresponding rodent complementarity determining regions. Accordingly, such humanised antibodies are chimeric antibodies, wherein substantially less than an intact human variable domain has been substituted by the corresponding sequence from a non-human species. In practice, humanised antibodies may be typically human antibodies in which some complementarity determining region residues and possibly some framework residues are substituted by residues from analogous sites in rodent antibodies.

Human antibodies can also be identified using various techniques known in the art, including phage display libraries (see, for example, Hoogenboom & Winter, 1991, Marks et al., 1991, Cole et al., 1985, Boemer et al., 1991, the disclosures of which are incorporated herein by reference).

In one embodiment, wherein the class of any one or more of the, or each, of said antibodies and/or antigen binding fragments is independently selected from group consisting of IgY, IgG, IgM, IgA, IgD, IgE, camelid and IgNAR, or modified versions thereof; preferably, IgY.

In a particular embodiment, the antibodies and/or antigen binding fragments with specificity for a particular antigen(s) may be IgY, whilst the antibodies and/or antigen binding fragments for a different antigen(s) may not be IgY ; for example, (a) one or more antibodies and/or antigen binding fragments with specificity to an ACE-2 antigen and/or (b) one or more antibodies and/or antigen binding fragments with specificity to an NRP-1 antigen are in the IgY antibody format; and/or (c) one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 Spike 1 RBD antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments; and/or (d) one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 SI antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments are in a non-IgY antibody format (such as IgG).

In one embodiment, any one or more of the, or each, antibody and/or antibody fragment is independently selected from group consisting of a monoclonal antibody preparation, a polyclonal antibody preparation, and a single -chain antibody format; preferably, a polyclonal antibody.

In a particularly preferred embodiment, the antibodies and/or antigen binding fragments with specificity to an ACE-2 antigen and/or antibodies and/or antigen binding fragments with specificity to an NRP- 1 antigen are independently further characterised by one or more (preferably all) of the characteristics selected from the group consisting of: i. it does not comprise a sequence that displays binding to the human Fc receptor; ii. it does not comprise a sequence that activates the human complement system; iii. it does not comprise a sequence that is specifically bound by protein A; and iv. it does not comprise a sequence that is specifically bound by protein G.

In a further preferred embodiment, the antibodies and/or antigen binding fragments with specificity to an ACE-2 antigen and/or antibodies and/or antigen binding fragments with specificity to an NRP-1 antigen and/or SARS-CoV-2 Spike 1 RBD antigen and/or SARS-CoV-2 SI antigen are independently further characterised by one or more (preferably all) of the characteristics selected from the group consisting of: i. it does not comprise a sequence that displays binding to the human Fc receptor; ii. it does not comprise a sequence that activates the human complement system; iii. it does not comprise a sequence that is specifically bound by protein A; and iv. it does not comprise a sequence that is specifically bound by protein G.

Preferably, the antibodies and/or antigen binding fragments do not: i. comprise a sequence that displays binding to the human Fc receptor; and/or ii. comprise a sequence that activates the human complement system.

In one embodiment, the antibodies and/or antigen binding fragments do not comprise a sequence that displays binding to the human Fc receptor: i. do not comprise an Fc region; ii. do not comprise one or more (preferably two) heavy chain constant domain 2 (CH2) and/or one or more (preferably two) heavy chain constant domain 3 (CH3); and/or, if present, one or more (preferably two) heavy chain constant domain 4 (CH4); or; iii. comprises an Fc region with one or more mutations that reduce and/or inhibit binding of the Fc region to the human Fc receptor.

In a particularly preferred embodiment, the composition (in particular, the antibodies and/or antigen binding fragments with specificity to an ACE-2 antigen and/or antibodies and/or antigen binding fragments with specificity to an NRP-1 antigen) does not promote an immune response in the subject.

In one embodiment, the immune response in the subject is characterised by antibody directed cytotoxicity (ADCC) mediated by the antibodies and/or antigen binding fragments with specificity to an ACE-2 antigen and/or antibodies and/or antigen binding fragments with specificity to an NRP-1 antigen. In a particular preferred embodiment, the antibodies and/or antigen binding fragments are one or more antibody and/or antibody binding fragment selected from the list consisting of: single domain antibodies (dAbs, including single and dual formats [i.e. dAb-linker-dAb], and nanobodies, preferably nanobodies); Fc-silent antibodies; and scFvs.

As described herein, the antibodies and/or antigen binding fragments may target the ACE-2 antigen and/or

NRP-1 antigen, which will be antigens endogenous within the subject (preferably human) to be treated. Accordingly, it is most preferable that the composition has no, or has a low risk, of leading to the subject’s immune response targeting those endogenous antigens. IgY is an example of a format of antibody that benefits from these characteristics. Additionally, as described further herein, IgY also has the advantage that it can be cost-effectively produced in chickens.

In one embodiment wherein the composition comprises two or more of (a), (b), (c) and/or (d), as discussed herein, the composition is more effective for the treatment and/or prevention of a coronavirus than a composition comprising only one of (a), (b), (c) and/or (d); preferably, there is a synergy.

In a second aspect of the further combination composition embodiments, the present invention provides a composition as defined in the first aspect of the invention for use in medicine (such as for use in the prevention and/or treatment of an infection of a coronavirus in a subject).

The term “treatment” would be understood by those skilled in medicine. The term "treatment" as used herein may include any treatment of a condition, (for example, wherein the condition is coronavirus), in a subject, particularly a human or other mammal, and optionally includes one or more of the following effects:

(i) inhibiting the condition (for example, coronavirus), i.e., slowing, reducing or arresting the development of the condition;

(ii) relieving the condition (for example, coronavirus), i.e., causing regression of the condition in a subject having the condition; or

(iii) curing the condition (for example, coronavirus), i.e. returning a subject having the condition, to a state of health in which the condition, is no longer detectable.

The term “prevention” would be understood by those skilled in medicine. The term "prevention" as used herein may include any prophylaxis or prophylactic treatment of a condition (for example, wherein the condition is coronavirus) in a subject, particularly a human or other mammal, and optionally includes one or more of the following effects:

(i) preventing the condition (for example, coronavirus) from occurring in a subject (for example, a subject which may be predisposed to, or at risk of, developing the condition, but has not yet been diagnosed as having it) i.e. stopping the subject from developing the condition - for example, the subject has been exposed to a coronavirus;

(ii) delaying the onset of the condition (for example, coronavirus) in a subject, i.e. delaying a subject from developing the condition - for example, delaying the subject from exhibiting the symptoms of coronavirus after exposure;

(iii) limiting the occurrence of the condition (for example, coronavirus) in a subject, for example reducing the extent to which a subject is affected by the condition; or

(iv) preventing one or more symptoms of the condition (for example, coronavirus) in a subject, i.e. stopping the subject from developing one or more symptoms of the condition.

A particular embodiment, the treatment and/or prevention can result in the reduction of one or more of the following characteristics:

• level of IL-6;

• extent and/or level of alveolitis;

• extent and/or level of alveolar damage;

• severity of alveolitis;

• extent and/or level of alveolitis and severity of alveolitis;

• extent and/or level of alveolar damage and severity of alveolitis;

• presence of an alveolar oedema;

• an alveolar haemorrhage;

• presence of a Type II pneumocyte hyperplasia;

• severity of bronchitis;

• severity of bronchiolitis;

• extent and/or level of peribronchial;

• extent and/or level perivascular cuffing; and/or

• severity of tracheitis.

Preferably, the treatment and/or prevention can result in the reduction of one or more of the following characteristics:

• severity of alveolitis; • extent and/or level of alveolitis and severity of alveolitis;

• extent and/or level of alveolar damage and severity of alveolitis;

• presence of a Type II pneumocyte hyperplasia; and/or

• severity of tracheitis.

The term “subject” as used herein includes the meaning of any living human or animal (humans and mammals are of particular interest) to which the treatment of the present invention is applied.

In one embodiment, the infection of a coronavirus can be characterised by one or more of the following:

• the subject has tested positive for coronavirus, but is yet to exhibit a symptom;

• the subject is exhibiting a mild coronavirus symptom;

• the subject is or has been or is at risk of being hospitalized due to the infection of a coronavirus;

• the subject is being treated with or has been treated with or is at risk of being treated with a ventilator due to the infection of a coronavirus; and/or

• the subject has or has had or is at risk of having sepsis due to the infection of a coronavirus;

• the subject has or has had or is at risk of having a cytokine storm due to the infection of a coronavirus, preferably:

• the subject has tested positive for coronavirus, but is yet to exhibit a symptom; and/or

• the subject is exhibiting mild coronavirus symptoms.

In one embodiment, the subject in unable to receive a coronavirus vaccine; for example, for medical reasons.

In an alternative second aspect of the further combination composition embodiments, the present invention provides a first composition (such as defined in the first aspect of the invention) for use in medicine (such as for use in the prevention and/or treatment of an infection of a coronavirus in a subject), wherein the composition comprises, consists essentially of, or consists of one or more (preferably, one to three) antibodies and/or antigen binding fragments, and/or one or more (preferably, one to three) nucleic acids encoding any one or more (preferably, one to three) of said antibodies and/or antigen binding fragments, selected from the group consisting of:

(a) one or more antibodies and/or antigen binding fragments with specificity to an ACE-2 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments;

(b) one or more antibodies and/or antigen binding fragments with specificity to an NRP-1 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments; (c) one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 Spike 1 RBD antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments; and

(d) one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 SI antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments, wherein the first composition is administered to a subject with at least a further composition comprising one or more antibodies and/or antigen binding fragments, and/or one or more nucleic acids encoding any one or more of said antibodies and/or antigen binding fragments, selected from the group consisting of, and preferably not included in the first composition:

(a) one or more antibodies and/or antigen binding fragments with specificity to an ACE-2 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments;

(b) one or more antibodies and/or antigen binding fragments with specificity to an NRP-1 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments;

(c) one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 Spike 1 RBD antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments; and

(d) one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 SI antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments, and/or the further composition is a further therapeutic agent.

In a third aspect of the further combination composition embodiments, the present invention provides a use of one or more antibodies and/or antigen binding fragments, and/or one or more nucleic acids encoding any one or more of said antibodies and/or antigen binding fragments, as defined in the first aspect of the invention in the manufacture of a medicament for the prevention and/or treatment of an infection of a coronavirus in a subject.

In an alternative third aspect of the further combination composition embodiments, the present invention provides a use of one or more (preferably, one to three) antibodies and/or antigen binding fragments, and/or one or more (preferably, one to three) nucleic acids encoding any one or more (preferably, one to three) of said antibodies and/or antigen binding fragments, (such as defined in the first aspect of the invention) in the manufacture of a first medicament for the prevention and/or treatment of an infection of a coronavirus in a subject, wherein the one or more antibodies and/or antigen binding fragments, and/or one or more nucleic acids encoding any one or more of said antibodies and/or antigen binding fragments, selected from the group consisting of:

(a) one or more antibodies and/or antigen binding fragments with specificity to an ACE-2 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments;

(b) one or more antibodies and/or antigen binding fragments with specificity to an NRP-1 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments;

(c) one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 Spike 1 RBD antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments; and

(d) one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 SI antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments, wherein the first medicament is administered to a subject with at least a further composition comprising one or more antibodies and/or antigen binding fragments, and/or one or more nucleic acids encoding any one or more of said antibodies and/or antigen binding fragments, selected from the group consisting of, and preferably not included in the first medicament:

(a) one or more antibodies and/or antigen binding fragments with specificity to an ACE-2 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments;

(b) one or more antibodies and/or antigen binding fragments with specificity to an NRP-1 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments;

(c) one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 Spike 1 RBD antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments; and

(d) one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 SI antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments, and/or the further composition is a further therapeutic agent.

In a fourth aspect of the further combination composition embodiments, the present invention provides a method for the prevention and/or treatment of an infection of a coronavirus in a subject, comprising administering to the subject a therapeutically effective amount of a composition as defined in the first aspect of the invention.

The term “therapeutically effective amount” refers to the amount of a drug, compound, protein (for example the compositions, antibodies and antigen-binding fragments as defined herein) or pharmaceutical composition that is sufficient to effect beneficial or desired results including clinical results associated with the treatment or prevention of coronavirus and/or a condition associated therewith. An effective amount can be administered in one or more administrations. For purposes of this invention, an effective amount of drug, compound, protein (for example the compositions, antibodies and antigen-binding fragments as defined herein), pharmaceutical composition or veterinary composition is an amount sufficient to treat, ameliorate, reduce the intensity of and/or prevent coronavirus and/or a condition associated therewith. As is understood in the clinical context, an effective amount of a drug, compound, protein (for example the compositions, antibodies and antigenbinding fragments as defined herein) or pharmaceutical composition may or may not be achieved when administered in conjunction (such as separately, sequentially or simultaneously) with another drug, compound, protein or pharmaceutical, such as those described elsewhere in this application. Thus, an “effective amount” may be considered in the context of administering one or more therapeutic agents, and a single agent may be considered to be given in an effective amount if, in conjunction with one or more other agents, a desirable result may be or is achieved.

In an alternative fourth aspect of the further combination composition embodiments, the present invention provides a method for the prevention and/or treatment of an infection of a coronavirus in a subject, comprising administering to the subject a first composition (such as defined in the first aspect of the invention) comprising, consisting essentially of, or consisting of one or more (preferably, one to three) antibodies and/or antigen binding fragments, and/or one or more (preferably, one to three) nucleic acids encoding any one or more (preferably, one to three) of said antibodies and/or antigen binding fragments, selected from the group consisting of:

(a) one or more antibodies and/or antigen binding fragments with specificity to an ACE-2 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments;

(b) one or more antibodies and/or antigen binding fragments with specificity to an NRP-1 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments; (c) one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 Spike 1 RBD antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments; and

(d) one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 SI antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments, wherein the first composition is administered to a subject with at least a therapeutically effective amount of a further composition comprising one or more antibodies and/or antigen binding fragments, and/or one or more nucleic acids encoding any one or more of said antibodies and/or antigen binding fragments, selected from the group consisting of, and preferably not included in the first composition:

(a) one or more antibodies and/or antigen binding fragments with specificity to an ACE-2 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments;

(b) one or more antibodies and/or antigen binding fragments with specificity to an NRP-1 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments;

(c) one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 Spike 1 RBD antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments; and

(d) one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 SI antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments, and/or the further composition is a further therapeutic agent.

It will be appreciated that any, and all of the features, of the first aspect of the invention are compatible with, and included in, the second to fourth aspects of the invention; in particular, the combinations of the antibodies and/or antigen binding fragments, and/or the nucleic acids encoding any one or more of said antibodies and/or antigen binding fragments described therein.

In a particular embodiment of the second to fourth aspects of the invention, the first composition comprises, consists essentially of, or consists of a form of antibodies and/or antigen binding fragments, and/or nucleic acids encoding said antibodies and/or antigen binding fragments, which is different to the form of antibodies and/or antigen binding fragments, and/or nucleic acids encoding said antibodies and/or antigen binding fragments in the further compositions. In one embodiment of the alternative second to fourth aspects of the invention, the first composition (or first medicament) is administered before, after and/or concurrently with the further composition.

In one embodiment of the alternative second to fourth aspects of the invention, the further composition is a second composition and a third composition, and optionally a fourth composition. In one embodiment, the first composition (or first medicament), second composition, third composition, and/or fourth composition are administered sequentially. It will appreciated that this sequential administration could be in any order.

One embodiment of the alternative second to fourth aspects of the invention includes a first composition (or medicament), a second composition, and a third composition and/or a fourth composition, wherein each composition comprises, consists essentially of, or consists of one of:

(a) one or more antibodies and/or antigen binding fragments with specificity to an ACE-2 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments;

(b) one or more antibodies and/or antigen binding fragments with specificity to an NRP-1 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments;

(c) one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 Spike 1 RBD antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments; or

(d) one or more antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 SI antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments, preferably, wherein each composition comprises antibodies and/or antigen binding fragments with specificity for a different antigen. It will be appreciated that each of the first composition (or medicament), a second composition, a third composition and a fourth composition may comprise one or more of the above (a) to (d).

In one embodiment of the alternative second to fourth aspects of the invention, there is a period of about 30 minutes or more from administration of the first composition (or first medicament) and the administration of the further composition(s); for example: about one hour or more; about two hours or more; about three hours or more; about four hours or more; about five hours or more; about six hours or more; about 12 hours or more; about 18 hours or more; about 24 hours or more; about two days or more; about three days or more; about four days or more; about five days or more; about six days or more; about seven days or more; about eight days or more; about nine days or more; about ten days or more; about 11 days or more; about 12 days or more; about 13 days or more; about 14 days or more; about three weeks or more; about four weeks or more; about two months or more; about three months or more; about four months or more; about five months or more; about six months or more; about seven months or more; about eight months or more; about nine months or more; about ten months or more; about 11 months or more; or about 1 year or more. In an alternative embodiment of the alternative second to fourth aspects of the invention, there is a period of about 30 minutes or less from administration of the first composition (or first medicament) and the administration of the further composition(s); for example: about one hour or less; about two hours or less; about three hours or less; about four hours or less; about five hours or less; about six hours or less; about 12 hours or less; about 18 hours or less; about 24 hours or less; about two days or less; about three days or less; about four days or less; about five days or less; about six days or less; about seven days or less; about eight days or less; about nine days or less; about ten days or less; about 11 days or less; about 12 days or less; about 13 days or less; about 14 days or less; about three weeks or less; about four weeks or less; about two months or less; about three months or less; about four months or less; about five months or less; about six months or less; about seven months or less; about eight months or less; about nine months or less; about ten months or less; about 11 months or less; or about 1 year or less. It will be appreciated that any ranges of periods described in the above two embodiments are encompassed herein, such as the period of 30 minutes to about 24 hours or the period of 30 minutes to about 1 year.

In one embodiment, the composition, or antibodies and/or antigen-binding fragments thereof, may be repeatedly administered (or the treatment regime of different compositions, or antibodies and/or antigen-binding fragments thereof, being administered at different times may be repeated). Of potentially particular relevance when a passive immunity is sought, the composition, or antibodies and/or antigen-binding fragments thereof, may be administered as often as necessary so that the antibodies for a specific antigen (ACE-2, NRP-1, RBD, and/or SI) are continuously present in the subject at a therapeutically effective level, for as long as the treatment is desired. Accordingly, the composition, or antibodies and/or antigen-binding fragments thereof, is again administered when the detectable antibodies and/or antigen-binding fragments thereof in the subject (for example, in the blood of the subject) are approaching a level at which the antibodies and/or antigen-binding fragments thereof may not be therapeutically effective.

Preferably, the further therapeutic agent is a therapeutic agent for the treatment and/or prevention of an infection of a coronavirus. In one embodiment of the alternative second to fourth aspects of the invention, the further therapeutic agent is a coronavirus vaccine, such as a coronavirus vaccine comprising SARS-CoV-2 Spike 1 RBD antigen and/or SARS-CoV-2 SI antigen, and/or a coronavirus vaccine comprising a nucleic acid encoding SARS-CoV-2 Spike 1 RBD antigen and/or SARS-CoV-2 SI antigen. Preferably in this embodiment, the first composition (or first medicament) comprises:

(a) one or more antibodies and/or antigen binding fragments with specificity to an ACE-2 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments; and/or

(b) one or more antibodies and/or antigen binding fragments with specificity to an NRP-1 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments.

In one embodiment, the coronavirus vaccine is a one or more coronavirus vaccine selected from the list consisting of: Comimaty; COVID- 19 Vaccine - Janssen; Spikevax (COVID- 19 Vaccine Modema); or Vaxzevria (CO VID-19 Vaccine AstraZeneca).

In one embodiment of the alternative second to fourth aspects of the invention, the further therapeutic agent is human serum from a subject that has previously been infected with a coronavirus; preferably, wherein the serum comprises detectable antibodies with specificity to the coronavirus. Preferably in this embodiment, the first composition (or first medicament) comprises:

(a) one or more antibodies and/or antigen binding fragments with specificity to an ACE-2 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments; and/or

(b) one or more antibodies and/or antigen binding fragments with specificity to an NRP-1 antigen and/or one or more nucleic acids encoding one or more of said antibodies and/or antigen binding fragments.

In one embodiment of the alternative second to fourth aspects of the invention, wherein the treatment and/or prevention comprises two or more of (a), (b), (c) and/or (d), as discussed herein, the treatment and/or prevention is more effective than the treatment and/or prevention when only one of (a), (b), (c) and/or (d) is administered to the subject; preferably, there is a synergy.

In one embodiment of the alternative second to fourth aspects of the invention in which the first composition (or first medicament) and further composition(s) comprise antibodies and/or antigen binding fragments with specificity for a different antigen(s), the combinations of antigens targeted are selected from the list consisting of: (a) and (b), as discussed herein; (a) and (c); (a) and (d); (b) and (c); (b) and (d); (c) and (d); (a), (b) and (c); (a), (b) and (d); (a), (c) and (d); (b), (c) and (d); and (a), (b), (c) and (d), preferably selected from the list consisting of: (a) and (c); (a) and (d); (a), (b) and (c), and (a), (b) and (d); and (a), (c) and (d), most preferably selected from: (a) and (c); (a), (b) and (c), and (a), (b) and (d), and even more preferably (a), (b) and (c) or (a) and (c).

In a preferred embodiment of the invention, the coronavirus is SARS-CoV-2.

In one embodiment of the invention, the coronavirus infection is a severe coronavirus infection.

In a particular embodiment, the severe coronavirus infection is characterised by elevated levels of IL- 6, preferably wherein the elevated levels of IL-6 are as described in the Examples.

The composition (or antibodies and/or antigen binding fragments described herein) may be, or may be subsequent formulated to form, a pharmaceutical or veterinary composition.

The final product may be presented in a unit dosage form. For example, a unit dosage form may contain about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 mg of the antibody and/or antigen binding fragment (wherein the term “about” refers to ± 0.5, 0.4, 0.3, 0.2 or 0.1 mg) or more, e.g. within the range of 0.1 to 1000 mg, or 1 to 100 mg.

A pharmaceutical or veterinary composition may comprise the antibodies and/or antigen binding fragments in admixture with a pharmaceutically or veterinary acceptable adjuvant, diluent or carrier, which will typically be selected with regard to the intended route of administration and standard pharmaceutical practice. The composition may be in the form of immediate-, delayed- or controlled- re lease applications. Preferably, the formulation is a unit dosage containing a daily dose or unit, daily sub-dose or an appropriate fraction thereof, of the active ingredient.

The phrases “pharmaceutical or veterinary acceptable” include reference to compositions that do not produce an adverse, allergic or other untoward reaction when administered to an animal or a human, as appropriate. The preparation of such pharmaceutical or veterinary compositions are known to those of skill in the art in light of the present disclosure, as exemplified by Remington's Pharmaceutical Sciences, 18th Ed. Mack Printing Company, 1990, incorporated herein by reference. Moreover, for animal or human administration, it will be understood that preparations should meet sterility, pyrogenicity, general safety and purity standards as required by FDA Office of Biological Standards. As used herein, “pharmaceutically or veterinarily acceptable carrier” includes any and all solvents, dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antibacterial agents, antifungal agents), isotonic agents, salts, preservatives, drugs, drug stabilizers, excipients, disintegration agents, such like materials and combinations thereof, as would be known to one of ordinary skill in the art. Except insofar as any conventional carrier is incompatible with the active ingredient, its use in the therapeutic or pharmaceutical compositions is contemplated.

The pharmaceutical or veterinary compositions according to the invention may, or may not, be intended for, and, thus formulated in a manner suitable for, parenteral, intravenous, intra-arterial, intraperitoneal, intra-muscular, intra-ocular, intra-cranial, intra-cerebrally, intra-osseously, intra-cerebroventricularly, intra-thecally, intra-nasal, aerosol (by metered-dose inhalers or nebulizer devices), or subcutaneous administration. In one preferred embodiment, the formulation if for aerosol administration and/or intranasal administration. Sterile injectable solutions may be prepared by incorporating the active compounds in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by sterilization. The pharmaceutical compositions may be best used in the form of a sterile aqueous solution which may contain other substances, for example, enough salts or glucose to make the solution isotonic with blood. The aqueous solutions may be suitably buffered (preferably to a pH of from 3 to 9), if necessary. The preparation of suitable pharmaceutical formulations under sterile conditions is readily accomplished by standard pharmaceutical techniques well-known to those skilled in the art.

The pharmaceutical or veterinary compositions according to the invention may alternatively be formulated in the form of a powder, such as a sterile powder, which may be a lyophilised powder.

In the medical uses described herein, the administration is one or more type of administration selected from the list consisting of: parenteral, intravenous, intra-arterial, intraperitoneal, intra-muscular, intraocular, intra-cranial, intra-cerebrally, intra-osseously, intra-cerebroventricularly, intra-thecally, intra- nasal, aerosol (by metered-dose inhalers or nebulizer devices) and subcutaneous administration. In one preferred embodiment, the administration is aerosol administration and/or intranasal administration and/or administration to the upper and/or lower respiratory tract.

A therapeutically effective amount of an antibodies and/or antigen binding fragments for administration to a subject, such as a human subject, on the basis of a daily dosage level may be from 0.01 to 1000 mg of antibodies and/or antigen binding fragments per adult (for example, from about 0.001 to 20 mg per kg of the subject’s body weight, such as 0.01 to 10 mg/kg, for example greater than 0.1 mg/kg and up to or less than 20, 10, 5, 4, 3 or 2 mg/kg, such as about 1 mg/kg), administered in single or divided doses.

The physician in any event will determine the actual dosage which will be most suitable for any individual subject and it will vary with the age, weight and response of the particular subject. The above dosages are exemplary of the average case. There can, of course, be individual instances where higher or lower dosage ranges are merited and such are within the scope of this invention.

For veterinary use, a compound of the invention is administered as a suitably acceptable formulation in accordance with normal veterinary practice and the veterinary surgeon will determine the dosing regimen and route of administration which will be most appropriate for a particular animal.

A further aspect of the invention provides a composition and/or antibodies and/or antigen binding fragments thereof, for the diagnosis of coronavirus. In particular, the antibodies and/or antigen binding fragments with specificity to RBD and/or SI are for particular interest for use in such diagnostic approaches. In such an embodiment, the antibodies and/or antigen binding fragments may be tagged with a reporter, such as a fluorescent reporter. Preferably, such a diagnosis is an in vitro method; for example, wherein a sample is provided from a subject, the composition and/or antibodies and/or antigen binding fragments thereof are applied to the sample, and wherein from the composition and/or antibodies and/or antigen binding fragments thereof being are applied to the sample the subject can be identified as being infected with coronavirus.

IgY antibody combination compositions

In aspect i, the present invention provides a composition comprising, consisting essentially of, or consisting of, at least two, three or four different forms of IgY antibodies and/or antibody fragments thereof, wherein said different forms are selected from the group consisting of:

(a) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to an Angiotensin Converting Enzyme 2 (ACE-2) antigen;

(b) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to a Neuropilin 1 (NRP-1) antigen;

(c) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen; and (d) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen, optionally wherein the two or more IgY antibodies and/or antibody fragments thereof are selected from the list consisting of: (a) and (b); (a) and (c); (a) and (d); (b) and (c); (b) and (d); (c) and (d); (a), (b) and (c); (a), (b) and (d); (a), (c) and (d); (b), (c) and (d); and (a), (b), (c) and (d), preferably selected from the list consisting of: (a) and (c); (a) and (d); (a), (b) and (c), and (a), (b) and (d); and (a), (c) and (d), most preferably selected from: (a) and (c); (a), (b) and (c), and (a), (b) and (d), and even more preferably (a), (b) and (c) or (a) and (c).

The ACE-2, NRP-1, RBD and/or SI antigens (and/or immunogenic fragments and/or immunogenic variants thereof) can be as described above for the “Antibody combination compositions” aspects.

In one embodiment, the composition is for passive immunization.

In one embodiment, the

(i) one or two forms of IgY antibodies and/or antibody fragments thereof as present in said composition comprise, consist essentially of, or consist of, one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to an ACE-2 antigen, and/or one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to an NRP-1 antigen, and

(ii) one or two forms of IgY antibodies and/or antibody fragments thereof present in said composition comprise, consist essentially of, or consist of, one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike 1 RBD antigen, and/or one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 S 1 antigen.

In one embodiment, the

(i) the forms of IgY antibodies or antibody fragments thereof as present in said composition comprise, consist essentially of, or consist of, one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to an ACE-2 antigen, and one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to an NRP-1 antigen, or

(ii) the forms of IgY antibodies or antibody fragments thereof as present in said composition comprise, consist essentially of, or consist of, one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike 1 RBD antigen, and one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 S 1 antigen. In one embodiment, the the forms of IgY antibodies or antibody fragments thereof as present in said composition comprise, consist essentially of, or consist of:

(i) one or more forms of IgY antibodies or antibody fragments thereof with specificity to an ACE-2 antigen, and one or more forms of IgY antibodies or antibody fragments thereof with specificity to SARS-CoV-2 Spike 1 RBD antigen,

(ii) one or more forms of IgY antibodies or antibody fragments thereof with specificity to an ACE-2 antigen, and one or more forms of IgY antibodies or antibody fragments thereof with specificity to SARS-CoV-2 SI antigen,

(iii) one or more forms of IgY antibodies or antibody fragments thereof with specificity to an NRP-1 antigen, and one or more forms of IgY antibodies or antibody fragments thereof with specificity to SARS-CoV-2 Spike 1 RBD antigen; or

(iv) one or more forms of IgY antibodies or antibody fragments thereof with specificity to a Neuropilin 1 (NRP-1) antigen, and one or more forms of IgY antibodies or antibody fragments thereof with specificity to SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

In a preferred embodiment, the the forms of IgY antibodies and/or antibody fragments thereof as present in said composition comprise, consist essentially of, or consist of:

- one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to an ACE-2 antigen;

- one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to an NRP-1 antigen; and

- one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike 1 RBD antigen.

In a more preferred embodiment, the the forms of IgY antibodies and/or antibody fragments thereof as present in said composition comprise, consist essentially of, or consist of:

- one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to an ACE-2 antigen; and

- one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike 1 RBD antigen.

In one embodiment, the one, two, three, or four, of the forms of IgY antibodies and/or antibody fragments thereof, as present in said composition, are polyclonal, and optionally wherein all of the forms of IgY antibodies and/or antibody fragments thereof, as present in said composition, are polyclonal. Preferably, the polyclonal IgY antibodies and/or antibody fragments thereof are monospecific.

One approach is to use egg-laying poultry, also referred to as layer, that is immunized, preferably with one or more antigens that are specific to the corona virus in question, and one or more antigens derived from the surface receptors ACE-2 and NRP-1 on human cells, and preferably to use the IgY antibody produced by the egg-laying poultry to prevent or treat a corona virus or other virus infections in an animal, preferably a human. Preferably, the immunization is made to stimulate production of IgY antibodies targeting the antigens.

The antibodies of the invention, preferably polyclonal antibodies, may be produced by immunization of egg-laying poultry with a single or a combination of any of the following antigens: Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-1) antigen, SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, and SARS-CoV-2 Spike Glycoprotein 1 (SI). The ACE-2, NRP-1, RBD and/or SI antigens as relevant to this embodiment may be those described in the “Antibody combination compositions” section of this application, including but not limited to any of the amino acid sequences SEQ ID NOs: 1-8, or immunogenic fragments and/or variants thereof.

The present invention relates to GMP (Good Manufacturing Practice) production and fractionation of SARS-CoV-2 (and other respiratory viruses), ACE-2 and NRP-1 specific polyclonal antibodies in layers. The egg-laying poultry (layers) used for the production of antibodies is preferably Gallus gallus domesticus. A formulation of the polyclonal antibody will preferably be produced, and the formulation will preferably be administered to animals, such as humans, prophylactically for preventing an infection or as therapy for the treatment of an ongoing disease by the disease-causing virus.

In one embodiment, the wherein one, two, three, or four, of the forms of the polyclonal IgY antibodies and/or antibody fragments thereof, as present in said composition, are obtained by, or obtainable by, by a method that comprises a step of immunization of an egg-lying poultry (layer) with one or more antigens selected from the group consisting of:

(a) an Angiotensin Converting Enzyme 2 (ACE-2) antigen;

(b) a Neuropilin 1 (NRP-1) antigen;

(c) a SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen; and

(d) a SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

In a preferred embodiment, the polyclonal IgY antibodies and/or antibody fragments thereof are obtained by, or obtainable by, immunization of an egg-lying poultry (layer) with: (a) an ACE-2 antigen;

(b) an NRP-1 antigen; and

(c) a SARS-CoV-2 Spike 1 RBD antigen.

In a more preferred embodiment, the polyclonal IgY antibodies and/or antibody fragments thereof are obtained by, or obtainable by, immunization of an egg-lying poultry (layer) with:

(a) an ACE-2 antigen; and

(c) a SARS-CoV-2 Spike 1 RBD antigen.

In one embodiment in which the composition comprises polyclonal IgY antibodies or antibody fragments thereof are obtained by, or obtainable by, immunization of an egg-lying poultry (layer), the polyclonal IgY antibodies or antibody fragments thereof are isolated from an egg from the egg-laying poultry; preferably, the polyclonal IgY antibodies or antibody fragments thereof are purified following being isolated.

The antigens relevant to these embodiments may be those discussed above in relation to the “Antibody combination compositions”.

In one embodiment, the IgY antibodies or antibody fragments are enriched.

In one embodiment, the IgY antibodies or antibody fragments are enriched using the corresponding antigen and/or an epitope from the corresponding antigen.

In one embodiment, the IgY antibodies or antibody fragments are purified and/or enriched by affinity capture.

In one embodiment, the composition is characterised by one or both of: i. the absence of or low detectable levels of non-antibody and/or non-antibody fragment egg protein; ii. the absence of or detectable levels of non-ACE-2, non-NRP-1, non-RBD, and/or non- SI IgY antibodies or antibody fragments, and/or high levels of ACE-2, NRP-1, RBD, and/or S 1 IgY antibodies or antibody fragments when compared to non-ACE-2, non- NRP-1, non-RBD, and/or non-Sl IgY antibodies or antibody fragments. In one embodiment in which the composition comprises polyclonal IgY antibodies and/or antibody fragments thereof are obtained by, or obtainable by, immunization of an egg-lying poultry (layer), each IgY antibody and/or antibody fragment thereof with specificity to a specific antigen is obtained from, or obtainable from, an egg-lying poultry that has been immunized only with the corresponding specific antigen, and wherein the composition comprises a mixture of IgY antibodies and/or antibody fragments obtained from, or obtainable from, the egg from each egg-laying poultry immunized.

In a preferred embodiment in which the composition comprises polyclonal IgY antibodies and/or antibody fragments thereof are obtained by, or obtainable by, immunization of an egg-lying poultry (layer), the IgY antibodies and/or antibody fragments thereof are obtained from, or obtainable from, a egg -lying poultry that has been immunized with each of the antigens for which the IgY antibodies and/or antibody fragments thereof have specificity. As discussed herein, this can be referred to as multiplex immunization. As discussed in the Examples, producing the IgY antibodies in this manner produces a composition that is particularly effective at preventing and/or treating a coronavirus.

The SARS-CoV-2, ACE-2 and NRP-1 polyclonal antibodies (IgY) of the present invention may be produced by immunization of the egg-laying poultry with one or more, e.g. a mixture of antigens, such as antigens specific for corona viruses combined with SARS-CoV-2 specific Spike antigens and HEK293, Pichia and/or E coli expressed ACE-2 and NRP- 1 protein antigens, respectively. Suitably, the immunization takes place by injection, preferably by intramuscular injection. The polyclonal IgY antibodies may then be extracted, isolated and purified from the egg yolk of the eggs.

Preferably, the IgY antibody composition according to the invention is obtained by immunization of an egg-laying poultry with a single or a combination of any of the following antigens: Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP- 1) antigen, SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, and SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

Preferably, the IgY antibody composition for use in prevention and/or treatment of an infection of a coronavirus in an animal according to the invention is obtained by immunization of an egg-laying poultry with a single or a combination of any of the following antigens: Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-1) antigen, SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, and SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

Preferably, the egg yolk from an egg from an egg-laying poultry (layer) for use in prevention and/or treatment of an infection of a coronavirus in an animal according to the invention comprises IgY antibodies obtained by immunization of the egg-laying poultry with a single or a combination of any of the following antigens: Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-l) antigen, SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, and SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

The IgY antibody composition and IgY antibodies referred to herein may be obtained by immunization of an egg-laying poultry (layer) by a method which comprises:

(i) immunization of an egg-laying poultry (layer) with one or more antigens as defined herein, preferably the immunization is made by providing or administering said one or more antigen in the form of an injection, or two or more injections,

(ii) collecting egg of the egg-laying poultry,

(iii) obtaining egg yolk from the egg,

(iv) extracting and isolating the IgY antibody composition and/or IgY antibodies from the egg yolk and purifying the IgY antibody composition and/or IgY antibodies.

The immunization of an egg-laying poultry (layer) referred to above preferably takes place by one or more intramuscular injections, e.g. every two to four weeks.

The egg-laying poultry of the invention is preferably selected from chicken, or Gallus gallus domesticus, including hens, e.g. from flocks of layers from a Norwegian egg-production facility.

The extraction and isolation of the IgY antibody composition and/or IgY antibodies from the egg yolk may take place by removal of fat-soluble components by dilution with water and salt precipitation, e.g. by using NaCl, of the IgY antibody composition and/or IgY antibodies to achieve a purity of 90-95%. If desired, further purification of the IgY antibodies and/or IgY antibody compositions may take place, e.g. by chromatographical techniques or other methods for antibody purifications.

In a preferred embodiment of the invention, the IgY antibody composition is obtained by immunization of an egg-laying poultry (layer) with an Angiotensin Converting Enzyme 2 (ACE-2) antigen.

In a preferred embodiment of the invention, the IgY antibody composition is obtained by immunization of an egg-laying poultry (layer) with Neuropilin 1 (NRP-l) antigen.

In a preferred embodiment of the invention, the IgY antibody composition is obtained by immunization of an egg-laying poultry (layer) with SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen. In a preferred embodiment of the invention, the IgY antibody composition is obtained by immunization of an egg-laying poultry (layer) with SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

In a preferred embodiment of the invention, the IgY antibody composition is obtained by immunization of an egg-laying poultry (layer) with two or more antigens selected from Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-l) antigen, SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen and SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen.

In situations where the IgY antibody composition is obtained by immunization of an egg-laying poultry (layer) with two or more antigens, preferred examples of such two or more antigens according to the invention include:

Angiotensin Converting Enzyme 2 (ACE-2) antigen and SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen,

Angiotensin Converting Enzyme 2 (ACE-2) antigen and SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen,

Angiotensin Converting Enzyme 2 (ACE-2) antigen and Neuropilin 1 (NRP-l) antigen, Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-l) antigen and SARS- CoV-2 Spike Glycoprotein 1 (SI) antigen,

Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-l) antigen and SARS- CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen,

Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-l) antigen, SARS- CoV-2 Spike Glycoprotein 1 (SI) antigen and SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, and

SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigens and SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

In aspect ii, the present invention provides a composition for use in medicine, wherein the composition is as defined herein.

In aspect iii, the present invention provides a composition for use in the treatment and/or prevention of an infection of a coronavirus in a subject, wherein the composition is as defined herein.

In aspect iv, the present invention provides a use of two or more IgY antibodies or antibody fragments thereof as defined herein in the manufacture of a medicament for the treatment and/or prevention of an infection of a coronavirus in a subject In aspect v, the present invention provides a method for the treatment and/or prevention of an infection of a coronavirus in a subject, comprising administering to the subject a therapeutically effective amount of a composition as defined herein.

In aspect vi, the present invention provides a first form of IgY antibody, and/or antibody fragment thereof, for use in medicine, wherein the first form of IgY antibody, and/or antibody fragment thereof, is selected from the group consisting of:

(a) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to an Angiotensin Converting Enzyme 2 (ACE-2) antigen;

(b) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to a Neuropilin 1 (NRP-1) antigen;

(c) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen; and

(d) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen, wherein the first form of IgY antibody and/or antibody fragment thereof is administered with one or more further forms of IgY antibody and/or antibody fragment, that differ from the first form (for example, do not have a specificity to the same antigen as the first IgY antibody or antibody fragment), wherein the one or more (for example, one, two or three) further forms of IgY antibody and/or antibody fragments is, or are, selected from the group consisting of:

(a) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to an Angiotensin Converting Enzyme 2 (ACE-2) antigen;

(b) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to a Neuropilin 1 (NRP-1) antigen;

(c) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen; and/or

(d) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen, and/or wherein the first IgY antibody and/or antibody fragment thereof is administered with a further therapeutic agent.

In aspect vii, the present invention provides a A first form of IgY antibody and/or antibody fragment thereof for use in the treatment and/or prevention of an infection of a coronavirus in a subject, wherein the first form of IgY antibody and/or antibody fragment thereof is selected from the group consisting of: (a) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to an Angiotensin Converting Enzyme 2 (ACE-2) antigen;

(b) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to a Neuropilin 1 (NRP-1) antigen;

(c) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen; and

(d) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen, wherein the first form of IgY antibody and/or antibody fragment thereof is administered with one or more (for example, one, two or three) further forms of IgY antibody and/or antibody fragment thereof, that differ from the first form (for example, do not have a specificity to the same antigen as the first IgY antibody or antibody fragment), wherein the one or more further forms of IgY antibody and/or antibody fragments thereof is, or are, selected from the group consisting of:

(a) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to an Angiotensin Converting Enzyme 2 (ACE-2) antigen;

(b) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to a Neuropilin 1 (NRP-1) antigen;

(c) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen; and/or

(d) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen, and/or wherein the first IgY antibody and/or antibody fragment thereof is administered with a further therapeutic agent.

In aspect viii, the present invention provides a use of a first form of IgY antibody and/or antibody fragment thereof in the manufacture of a medicament for the treatment and/or prevention of an infection of a coronavirus in a subject, wherein the first form of IgY antibody and/or antibody fragment thereof is selected from the group consisting of:

(a) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to an Angiotensin Converting Enzyme 2 (ACE-2) antigen;

(b) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to a Neuropilin 1 (NRP-1) antigen;

(c) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen; and (d) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen, wherein the medicament is administered with one or more (for example, two, three or four) further forms of IgY antibody and/or antibody fragment thereof, that differ from the first form (for example, that do not have a specificity to the same antigen as the first IgY antibody or antibody fragment), wherein the one or more further forms of IgY antibody and/or antibody fragments is selected from the group consisting of:

(a) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to an Angiotensin Converting Enzyme 2 (ACE-2) antigen;

(b) one or more forms of Igy antibodies and/or antibody fragments thereof with specificity to a Neuropilin 1 (NRP-1) antigen;

(c) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen; and

(d) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen, and/or wherein the first IgY antibody and/or antibody fragment thereof is administered with a further therapeutic agent.

In aspect ix, the present invention provides a method for the treatment and/or prevention of an infection of a coronavirus in a subject, comprising administering to the subject a therapeutically effective amount of a a first form of IgY antibody and/or antibody fragment thereof selected from the group consisting of:

(a) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to an Angiotensin Converting Enzyme 2 (ACE-2) antigen;

(b) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to a Neuropilin 1 (NRP-1) antigen;

(c) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen; and

(d) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen, wherein the first form of IgY antibody and/or antibody fragment thereof is administered with therapeutically effective amount of one or more (for example, one, two, or three) further forms of IgY antibody and/or antibody fragment thereof, that differ from the first form (for example do not have a specificity to the same antigen as the first IgY antibody or antibody fragment), wherein the one or more further forms of IgY antibody and/or antibody fragments thereof is, or are, selected from the group consisting of:

(a) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to an Angiotensin Converting Enzyme 2 (ACE-2) antigen;

(b) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to a Neuropilin 1 (NRP-1) antigen;

(c) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen; and

(d) one or more forms of IgY antibodies and/or antibody fragments thereof with specificity to SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen, and/or wherein the first IgY antibody and/or antibody fragment thereof is administered with a further therapeutic agent.

In one embodiment, the first IgY antibody and/or antibody fragment thereof is administered before, after and/or concurrently with the further IgY antibodies and/or antibody fragments thereof.

In one embodiment of aspects vi-ix of the invention, the further forms of IgY antibody and/or antibody fragment thereof is a form of second IgY antibody and/or antibody fragment thereof and a form of third IgY antibody and/or antibody fragment thereof, and optionally a form of fourth IgY antibody and/or antibody fragment thereof. In one embodiment, the first form of IgY antibody and/or antibody fragment thereof, second form of IgY antibody and/or antibody fragment thereof, third form of IgY antibody and/or antibody fragment thereof, and/or fourth form of IgY antibody and/or antibody fragment thereof are administered sequentially. It will appreciated that this sequential administration could be the first form of, second form of, third form of and/or fourth form of IgY antibody and/or antibody fragment thereof being administered in any order.

One embodiment of aspects vi-ix of the invention includes a first form of IgY antibody and/or antibody fragment thereof, a second form of IgY antibody and/or antibody fragment thereof, and a third form of IgY antibody and/or antibody fragment thereof and/or a fourth form of IgY antibody and/or antibody fragment thereof, wherein each IgY antibody and/or antibody fragment thereof comprises one of:

(a) one or more forms of IgY antibodies and/or antigen binding fragments with specificity to an ACE-2 antigen;

(b) one or more forms of IgY antibodies and/or antigen binding fragments with specificity to an NRP-1 antigen; (c) one or more forms of IgY antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 Spike 1 RBD antigen; or

(d) one or more forms of IgY antibodies and/or antigen binding fragments with specificity to SARS-CoV-2 SI antigen.

In one embodiment of aspects vi-ix of the invention, there is a period of about 30 minutes or more from administration of the first form of IgY antibody and/or antibody fragment thereof and the administration of the further forms of IgY antibody(ies) and/or antibody fragment(s) thereof; for example: about one hour or more; about two hours or more; about three hours or more; about four hours or more; about five hours or more; about six hours or more; about 12 hours or more; about 18 hours or more; about 24 hours or more; about two days or more; about three days or more; about four days or more; about five days or more; about six days or more; about seven days or more; about eight days or more; about nine days or more; about ten days or more; about 11 days or more; about 12 days or more; about 13 days or more; about 14 days or more; about three weeks or more; about four weeks or more; about two months or more; about three months or more; about four months or more; about five months or more; about six months or more; about seven months or more; about eight months or more; about nine months or more; about ten months or more; about 11 months or more; or about 1 year or more. In an alternative embodiment of aspects vi-ix of the invention, there is a period of about 30 minutes or less from administration of the first form of IgY antibody and/or antibody fragment thereof and the administration of the further forms of IgY antibody(ies) and/or antibody fragment(s) thereof; for example: about one hour or less; about two hours or less; about three hours or less; about four hours or less; about five hours or less; about six hours or less; about 12 hours or less; about 18 hours or less; about 24 hours or less; about two days or less; about three days or less; about four days or less; about five days or less; about six days or less; about seven days or less; about eight days or less; about nine days or less; about ten days or less; about 11 days or less; about 12 days or less; about 13 days or less; about 14 days or less; about three weeks or less; about four weeks or less; about two months or less; about three months or less; about four months or less; about five months or less; about six months or less; about seven months or less; about eight months or less; about nine months or less; about ten months or less; about 11 months or less; or about 1 year or less. It will be appreciated that any ranges of periods described in the above two embodiments are encompassed herein, such as the period of 30 minutes to about 24 hours or the period of 30 minutes to about 1 year.

Preferably, the further therapeutic agent is a therapeutic agent defined above in respect of “Antibody combination compositions”. It will be appreciated that any, and all of the features, of any of the aspects and/or embodiments of the invention as described herein, and in particular in the section “Antibody combination compositions ” are compatible with, and included in, aspects i-ix of the invention - including the definitions (such as “treatment” and “prevention”, as well as the compositions described therein).

In a preferred embodiment of the invention, the IgY antibody composition for the use in prevention and/or treatment of an infection of a coronavirus in an animal is obtained by immunization of an egg- laying poultry with an Angiotensin Converting Enzyme 2 (ACE-2) antigen.

In a preferred embodiment of the invention, the IgY antibody composition for the use in prevention and/or treatment of an infection of a coronavirus in an animal is obtained by immunization of an egg- laying poultry with Neuropilin 1 (NRP-l) antigen.

In a preferred embodiment of the invention, the IgY antibody composition for the use in prevention and/or treatment of an infection of a coronavirus in an animal is obtained by immunization of an egg- laying poultry with SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen.

In a preferred embodiment of the invention, the IgY antibody composition for the use in prevention and/or treatment of an infection of a coronavirus in an animal is obtained by immunization of an egg- laying poultry with SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

In a preferred embodiment of the invention, the IgY antibody composition for the use in prevention and/or treatment of an infection of a coronavirus in an animal is obtained by immunization of an egg- laying poultry with two of more antigens selected from an Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-l) antigen, SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, and SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

In situations where the IgY antibody composition for the use in prevention and/or treatment of an infection of a coronavirus in an animal is obtained by immunization of an egg-laying poultry (layer) with two or more antigens, optional examples of such two or more antigens according to the invention include:

Angiotensin Converting Enzyme 2 (ACE-2) antigen and SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen,

Angiotensin Converting Enzyme 2 (ACE-2) antigen and SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen, Angiotensin Converting Enzyme 2 (ACE-2) antigen and Neuropilin 1 (NRP-l) antigen, Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-l) antigen and SARS- CoV-2 Spike Glycoprotein 1 (SI) antigen,

Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-l) antigen and SARS- CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen,

Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-l) antigen, SARS- CoV-2 Spike Glycoprotein 1 (SI) antigen and SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, and

SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen and SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

In a preferred embodiment of the invention, the egg yolk from an egg from an egg-laying poultry (layer) for use in prevention and/or treatment of an infection of a coronavirus in an animal comprises IgY antibodies obtained by immunization of the egg-laying poultry with an Angiotensin Converting Enzyme 2 (ACE-2) antigen.

In a preferred embodiment of the invention, the egg yolk from an egg from an egg-laying poultry (layer) for use in prevention and/or treatment of an infection of a coronavirus in an animal comprises IgY antibodies obtained by immunization of the egg-laying poultry with a SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

In a preferred embodiment of the invention, the egg yolk from an egg from an egg-laying poultry (layer) for use in prevention and/or treatment of an infection of a coronavirus in an animal comprises IgY antibodies obtained by immunization of the egg-laying poultry with Neuropilin 1 (NRP-l) antigen.

In a preferred embodiment of the invention, the egg yolk from an egg from an egg-laying poultry (layer) for use in prevention and/or treatment of an infection of a coronavirus in an animal comprises IgY antibodies obtained by immunization of the egg-laying poultry with SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen.

In a preferred embodiment of the invention, the egg yolk from an egg from an egg-laying poultry (layer) for use in prevention and/or treatment of an infection of a coronavirus in an animal, wherein the egg yolk comprises IgY antibodies obtained by immunization of the egg-laying poultry with two or more antigens selected from Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-l) antigen, SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, and SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen. In situations where the egg yolk comprises IgY antibodies obtained by immunization of the egg-laying poultry with two or more antigens, preferred examples of such two or more antigens according to the invention include:

Angiotensin Converting Enzyme 2 (ACE-2) antigen and SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen,

Angiotensin Converting Enzyme 2 (ACE-2) antigen and SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen,

Angiotensin Converting Enzyme 2 (ACE-2) antigen and Neuropilin 1 (NRP-l) antigen, Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-l) antigen and SARS- CoV-2 Spike Glycoprotein 1 (SI) antigen,

Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-l) antigen and SARS- CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen,

Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-l) antigen, SARS- CoV-2 Spike Glycoprotein 1 (SI) antigen and SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, and

SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen and SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

When using two or more antigens according to the invention, the immunization may take place as a multiplex immunization, in which the layer is immunized with multiple or all antigens at the same time one or more times, or as a separate immunization, in which the layer is immunized separately with one or more of said two or more antigens one or more times.

Preferably, the IgY antibody composition may comprises antibodies that are able to bind to both epitopes of the corona virus and receptors on the host cell the virus uses for binding and entry, either by immunization of individuals by a combination of antigens or by combining antibody batches that are purified from single-antigen-immunized egg laying poultry.

Preferably, the IgY antibody composition comprises IgY antibodies which are polyclonal.

The immunization of egg-laying poultry according to the invention may be conducted by providing the antigen, or combination of two or more antigens, as an injection, or two or more injections.

Preferably, the antigen specific for corona viruses is one or more antigens, e.g. a mixture of two or more antigens, that is/are specific for corona 3 viruses. Preferably, the Angiotensin Converting Enzyme 2 (ACE-2) antigen is expressed in HEK293 cells or the ACE-2 protein, available as Cat. 90211-C02H from SinoBiological.

Preferably, the SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen is recombinant SARS- CoV-2 Spike SI RBD (aa 319-541), expressed in HEK293 cells with His-tag.

Preferably, the Neuropilin 1 (NRP-l) antigen is the sequence Phe 22 - Lys 644 expressed in HEK293, available from 2bScientific.

Examples of preferred Spike antigens include the SARS-CoV-2 Spike Glycoprotein (SI (seql-674)) SHFc-Tag (HEK293).

In preferred embodiments of these aspects of the invention, the animal is a human. The IgY antibody composition, or antibodies, may be administered to the animal, or human, as an injection (intravenous, intramuscular, or subcutaneous); as a nasopharyngeal spray or drops, as an oral spray, drops, lozenger or mouthwash; or as an aerosol (by metered-dose inhalers or nebulizer devices) for reaching lower respiratory tracts. Further, in preferred embodiments of these aspects of the invention, the infection and/or disease caused by the corona virus is COVID- 19.

Preferably, the IgY antibody composition may comprises several epitope specific antibodies which are able to bind and block several sites that the corona virus is dependent on for host cell entry and infection.

In a preferred embodiment of the invention, the virus is SARS-CoV-2.

It is contemplated that any method or composition described herein can be implemented with respect to any other method or composition described herein. Preferences and options for a given aspect, feature or parameter of the invention should, unless the context indicates otherwise, be regarded as having been disclosed in combination with any and all preferences and options for all other aspects, features and parameters of the invention. For example, any of the compositions and/or antibodies and/or antigen binding fragments with specificity to each and any of the antigens ACE-2, NRP-l, RBD and/or SI described herein can be combined.

These, and further, aspects of the present invention will be better appreciated and understood when considered in conjunction with the following description and the accompanying drawings. It should be understood, however, that the following description, while indicating various aspects and embodiments of the invention and numerous specific details thereof, is given by way of illustration and not of limitation. Many substitutions, modifications, additions and/or rearrangements may be made within the scope of the invention without departing from the spirit thereof, and the invention includes all such substitutions, modifications, additions and/or rearrangements.

The use of the word “a” or “an” when used in conjunction with the term “comprising” in the claims and/or the specification may mean “one,” but it is also consistent with the meaning of “one or more,” “at least one,” and “one or more than one.”

The terms "optionally", “preferably”, “such as”, “for example” and the like denote that the subsequently described feature, characteristic, event or circumstances may or may not occur, and that the description includes instances where said feature, characteristic, event or circumstances occurs and instances in which it does not occur. For the avoidance of doubt, it is to be understood that, in some embodiments of the invention, any feature, characteristic, event or circumstance described in the context of "optionally", “preferably”, “such as”, “for example” and the like may be disclaimed from the scope of the claims.

It will be appreciated that the term “comprising” may encompass the terms “consisting essentially of’ and “consisting of’. In accordance with the disclosure herein, embodiments of the invention that are defined with reference to the term “comprising”, “comprise” or comprises” or equivalents thereof, can be considered to optionally refer to “consisting essentially of’, “consist essentially of’ or “consists essentially of’ or an equivalent thereof, respectively. Further, in accordance with the disclosure herein, embodiments of the invention that are defined with reference to the term “comprising”, “comprise” or comprises” or equivalents thereof, can be considered to optionally refer to “consisting”, “consist” or “consists” or an equivalent thereof, respectively.

The listing or discussion of an apparently prior-published document in this specification should not necessarily be taken as an acknowledgement that the document is part of the state of the art or is common general knowledge.

Example 1 The following general procedure was used throughout the examples for the production and testing of neutralizing polyclonal IgY antibodies targeting SARS-CoV-2 spike 1, SARS-CoV-2 SI RBD, human ACE-2 and human NRP-1, unless otherwise described:

The example describes the production of polyclonal antibodies, by immunization with commercially available virus antigens, that can be applied for prophylaxis, therapy and to reduce symptoms of diseases caused by pathogenic viruses.

For reasons of simplicity, the IgY antibody composition and IgY antibodies, including polyclonal antibodies, may be referred to in the examples and figures as IgY.

The following antigens were used to prepare the IgY antibody compositions:

1. SARS-CoV-2 Spike Glycoprotein 1 (SI) o Sequence 1-674 o Host HEK293 (Human embryonic kidney cells) o Conjugate Sheep Fc-Tag o Purchased from the Native Antigen Company

2. SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) o Sequence 1-223 o Host HEK293 o Strain Wuhan-Hu- 1 o Conjugate 6xHis-tag, C-terminus o Purchased from the Native Antigen Company

3. Angiotensin Converting Enzyme 2 (ACE-2) o Expressed in HEK293 cell o Obtained from NTNU Norway, Prof. Magnar Bjoras o Sequence: Glnl8-Ser740 o Host: HEK293 o Conjugate: His-tag

4. Neuropilin 1 (NRP-1) o Sequence Phe 22 - Lys 644 o Host HEK293 o Conjugate His Tag o Purchased from 2bScientific SARS-CoV-2 Spike 1 and Spike 1 RBD proteins bind to host cells animal ACE-2 receptors upon infection and by that gain entrance into target cells. In addition, NRP-1 on human cells are found to be used by SARS-CoV-2 as a route for infection.

Immunization

Intramuscular immunization (i.m.) (injection): Two immunizations of the egg-laying poultry (Gallus gallus domesticus) were performed 14 days apart and then every four weeks. Intramuscular injections in the breast muscle were performed with SI, RBD, ACE-2 and NRP-1, either separately or in different combinations, using Freund’s Complete Adjuvant:

The egg laying poultry are moved to research facilities upon laying eggs (23-25 weeks) 10-15 egg laying poultry are housed in each cage

The egg laying poultry are tested free of common viruses, and have no known disease at the start of the antibody production protocol

At start of the antibody production protocol the egg laying poultry have received two regulatory mandatory vaccines - against Marek’s disease (dayold chicks) and against coccidiosis (1 day old or before 3 weeks of age)

Different immunization protocols are initially applied to the cages to produce several batches to be evaluated for their efficiency towards SARS-CoV-2 in humans

All antigens are given in a dose of 1-10 pg/hen, optionally more than 10 pg/hen, together with Freund's Complete Adjuvant (i.m. injection) or 1-10 pg/hen, optionally more than 10 pg/hen, together with adjuvant MONTANIDE IMS 1313 VG N PR. Fractionation of IgY antibodies

Fractionation of IgY antibodies can be performed by standard methods, e.g. a method established by Norimun AS. In this example 1, the protocol for fractionation of IgY antibodies is as follows: a. Collection of yolk from the eggs of the egg-laying hens b. Removal of yolk lipids i. 7x dilution with distilled water, pH adjustment to 5 by 1 N HC1 at room temperature. ii. Freeze and thawing iii. Manual paper fdtration of thawed yolk mass to remove fatty acids and lipids. c. Salting out the IgY antibodies i. Addition of NaCl to 8.8 % w/w of filtrate of yolk mass. pH adjustment to 4 by

1 N HC1 at room temperature. ii. Stirring 2 hours at room temperature. iii. Centrifugation at 3800 g, 20 min at 4°C. iv. Pellet contains IgY antibodies which is collected manually or diluted in buffer. d. Optional freeze-drying of IgY antibody containing pellet to powder i. At 60 °C

ELISA assays

The protocol for the ELISA assays to determine the binding capacity of the produced IgY antibodies against the various antigens is as follows: a. Dilute the antigen composition to a desired concentration, usually 1 pg/ml, in PBS b. For coating 100 pl antigen solution is transferred to each well in a 96-well ELISA- plate, seal the plate and incubate at 4°C overnight c. Empty the plate and wash each well three times, using an ELISA washer, with 200 pl PBS containing 0,1% tween and 0,1% BSA d. For blocking, add 200 pl PBS-0,1 % tween 1 % BSA to each well and incubate for 1 hour at room temperature e. Measure protein concentration in IgY-samples and dilute so all the samples to be analyzed and compared have the exact same concentration of IgY f. Empty the plate and wash each well three times, using an ELISA washer, with 200 pl PBS containing 0,1% tween and 0,1% BSA g. Make dilution-series of the IgY-samples, for example 1: 100 - 1:218 700, a three-fold dilution for each step, in PBS-0,1% tween 0,1 % BSA. Add 100 pl to each well and incubate for 1 hour at room temperature h. Empty the plate and wash each well three times, using an ELISA washer, with 200 pl PBS containing 0,1% tween and 0,1% BSA i. Dilute the secondary antibody, HRP conjugated goat-anti-chicken IgY, according to manufacturer’s recommendations in PBS-0,1% tween 0,1 % BSA. Add 100 pl to each well and incubate for 1 hour at room temperature j . Empty the plate and wash each well three times, using an ELISA washer, with 200 pl PBS containing 0,1% tween and 0,1% BSA k. Prepare TMB solution following the manufacturer instructions and add 100 pl to each well. Measure absorption at 650 nm

Virus neutralization

Virus neutralization assays can be performed by standard protocols, e.g. protocols established by Norwegian University of Science and Technology (NTNU). In this example 1, the protocol for the virus neutralization assay is as follows: a. Approximately 4 x 10 ⁴ Vero-E6 cells were seeded per well in 96-well plates. b. The cells were grown for 24 h in DMEM supplemented with 10% FBS and Pen-Strep. c. IgY sample protein concentrations were quantified using NanoDrop and Bradford assay. d. IgY samples were prepared in 3-fold dilutions at 7 different concentrations in the virus growth medium (VGM) containing 0.2% BSA and Pen-Strep in DMEM. e. Virus SARS-CoV-2 isolated from hospitalized patients (hCoV-19/Norway/- Trondheim-E9/2020) was added to the samples to achieve a mol of 0.1 and incubated for Ih at 37 °C. f. 0,1 % DMSO was added to the control wells. g. The Vero-E6 cells were incubated for 72 h with VGM. h. After the incubation period, the medium was removed, and a CellTiter-Glo assay was performed to measure viability. In vivo neutralization

The IgY antibodies that have shown a neutralizing effect on the virus samples, and especially on SARS- CoV-2 virus, will be examined for an effect in animals, e.g. humans. The IgY antibodies may be administered to an animal, e.g. a human, by means of an injection, nose or oral product, or by inhalation. The antibodies can be used as a preventive measure, i.e. to prevent the virus from entering the host cells, to lessen symptoms or to treat an infection of the virus by neutralizing the effect of the virus by binding or complexing to the virus antigens used to provoke the antigen response.

Example 2

This example illustrates an ELISA analysis on IgY antibodies targeting SI and RBD.

The general procedure of example 1 was followed, unless otherwise described below.

As RBD is a domain within SI, it is expected that anti-RBD-IgY will bind to SI and anti-Sl-IgY will bind to RBD.

As shown in Figure 1, a (above) and b (below), the ELISA analysis indeed shows that IgY antibodies generated by immunization with either SI or RBD bind to their respective antigens. In Figure 1, two types of controls are included; NP represents antibody purified from egg-laying poultry before they were immunized, and Secondary ab (Sec ab) represents the background noise in the ELISA assay of the HRP-conjugated goat-anti-chicken secondary antibody.

Example 3

This example illustrates an ELISA analysis with virus lysate as an antigen.

The general procedure of example 1 was repeated, unless otherwise described below.

To investigate if the IgY antibody binds to the virus, a virus lysate from authentic SARS-CoV-2 was used. The virus lysate contains all the virus proteins, including the spike protein. If the IgY antibody, produced via immunization of egg-laying poultry with recombinant proteins produced in HEK cells, binds to a virus lysate, it strongly indicates that the IgY antibody also will recognize living virus particles.

As shown in Figure 2, a (above) and b (below), the ELISA analysis shows that anti-Sl-IgY and anti- RBD-IgY recognize and bind to a virus lysate. The combination IgY sample (IgY harvested from eggs from layers immunized with SI, RBD, ACE-2 and NRP-1) (designated Kombo) shows the highest activity against the virus lysate. Anti-Sl-IgY and anti-RBD-IgY shows equal activity against the virus lysate. NP and Sec ab have the same meaning as in example 2.

Example 4

This example illustrates an ELISA analysis on IgY targeting NRP-1 and ACE-2.

The general procedure of example 1 was repeated, unless otherwise described below.

10 layers were immunized with either ACE-2 antigen, NRP-1 antigen or all the four antigens, i.e. a combination of ACE-2 + NRP-1 + SI + RBD (designated Kombo). After a series of immunizations over a few weeks the layers start to produce IgY antibodies towards the respective antigen(s). The IgY antibodies were harvested from the egg yolks. 20 ng of antigen were coated in each well. The IgY antibodies were tested in a concentration range between 0,23-500 pg/ml (dilution series). The development of color was measured by a spectrophotometer (light absorption at 650 nm).

As shown in Figure 3, a (above) and b (below), the layers immunized with NRP-1 and Kombo produce antibodies that recognize NRP-1, the layers immunized with ACE-2 and Kombo produce antibodies that recognize ACE-2, and finally the Kombo sample contains IgY antibodies that recognizes, with high specificity, both NRP-1 and ACE-2 antigens. NP and Sec ab have the same meaning as in example 2.

Example 5

This example illustrates an in vitro cell-based neutralization assay using authentic SARS-CoV-2 virus.

The general procedure of example 1 was followed, unless otherwise described below. For a relatively fast analysis on whether antibodies, or other kinds of therapy, holds the capacity to inhibit a virus, an in vitro cell-based neutralization assay is performed. A relevant cell culture is plated in a 96-well plate. For SARS-CoV-2 a relevant cell line would be one that express ACE-2 and NRP-1 on the cell surface. This would allow SARS-CoV-2 to infect the cells. A cell line that express both ACE-2 and NRP-1 is VERO E6 cells.

The protocol is such that first for an hour, the antibodies of interest is incubated (mixed) with authentic SARS-CoV-2. By doing so, antibodies have direct access to the virus and under these ideal conditions, if the IgY recognize the virus, the antibodies will bind. If the IgY samples contain antibodies that recognize strategic epitopes (regions) on either SI or RBD, they will bind and block essential sites for the virus. When these sites are blocked the virus is unable to infect cells; they are neutralized. After an hour, the IgY/SARS-CoV-2 mix is transferred to the cell cultures and incubated for another 72 hrs. This gives the virus enough time to infect the cells, if they are able to do so. If after 72 hrs the virus are unable to infect, it means that the IgY antibodies have neutralized the virus and protected the cells from being infected. A dilution series of the IgY antibodies are being tested in order to see how much the antibodies can be diluted and at the same time keep above 50% efficiency.

As shown in Figure 4, when anti-RBD-IgY and anti-ACE2-IgY are mixed (manually, they are not harvested from the same egg), a synergistic effect is observed. Together, these two types of IgY have a more potent neutralizing effect compared to either alone. The actual effect realised is likely greater than that visualised within the corresponding graph because manner in which the antibodies in the RBD and ACE-2 treatment are diluted leads to it containing half the amount of each of RBD IgY and ACE- 2 IgY when compared to the single treatments.

Example 6

This example shows the effect of the different IgY combinations.

The general procedure of example 1 was followed, unless otherwise described below.

NRP-1 + SI and RBD + ACE-2 are IgY solutions manually mixed in the laboratory, i.e. the individual types of IgY were extracted from separate eggs. In contrast, the designation “Kombo” is from layers immunized with all four antigens, i.e. ACE-2 + NRP-1 + RBD + SI; thus, four different types of specific IgY harvested from the same egg - this can be referred to as multipleximmunization. Multipleximmunization: All four antigens are mixed and injected simultaneously. When IgY is extracted they are isolated from the same egg. For multiplex-immunization with four antigens: 4 x 10 pg antigen per immunization per hen (10 pg of each). When immunized with only one antigen: 10 pg antigen per immunization per hen.

Kombo M.M: An IgY-sample where the four types of IgY are manually mixed at the lab (2,5 ml from each of the four, with the concentration of 10 mg/ml, are mixed to obtain 10 ml of a 10 mg/ml solution)

As shown in Figure 5, a (above), the Kombo sample increased the neutralizing effect.

Similarly, as shown in Figure 5, b (below), it is evident that it is beneficial to combine anti-RBD-IgY with anti -ACE2 -IgY, and to immunize layers with multiple antigens.

Example 7

Building on the work in Example 6, Example 7 includes further comparisons of the potency of the antibody compositions based on the manner in which they have been produced, by immunization of layers.

As shown in Figure 6, relates to cell viability, the Kombo composition (i.e. the layer immunized with all four antigens, i.e. ACE-2 + NRP-1 + RBD + SI) is more effective than the Kombo M.M. composition, in which the IgY are extracted from layers immunized by different antigens and then manually mixed. This further shows that it is beneficial to immunize a layer with multiple antigen. The x-axis shows the dilution factor. The starting concentration is 10 mg/ml.

As described below, Figure 7 relates to a neutralisation assay using the mCherry reporter. The method used is as follows. Approximately 4 x 10 ⁴ Vero-E6 cells (Merck) were seeded per well in 96-well plates. The cells were grown for 24 h in DMEM supplemented with 10% FBS and Pen-Strep. Protein concentration of the IgY samples were quantified using NanoDrop. IgY samples were prepared in 3- fold dilutions at 7 different concentrations, in the virus growth medium (VGM) containing 0.2% BSA and Pen-Strep in DMEM. Virus hCoV-19/Norway/Trondheim-E9/2020 was added to the samples to achieve a moi of 0. 1 and incubated for Ih at 37 °C. 0,1 % DMSO was added to the control wells. The preincubation of the virus with the IgY allows the viral antigen-specific antibodies to bind to the viral antigens ahead of being added to cells, whilst the host cell-specific antibodies are not able to access their antigen until the preincubation mixture is added to the cells. The Vero-E6 cells were incubated for 72 h with VGM. After the incubation period, the medium was removed, and a CellTiter-Glo assay was performed to measure viability. To measure level of infection 96-well plates were scanned in a plate reader for total mCherry-fluorescense per well (hCoV-19/Norway/Trondheim-E9/2020 express m Cherry).

As shown in Figure 7, using an mCherry assay, the Kombo composition and Kombo M.M. composition appears to have a similar effect in relation to inhibiting cell infection with the virus. The x-axis shows the dilution factor. The starting concentration is 10 mg/ml.

Example 8

This example illustrates a neutralization assay in terms of percent infection.

The general procedure of example 1 was followed, unless otherwise described below.

Percent infection was measured by making the virus fluorescent. The virus was made fluorescent by expression of a gene coding for mCherry and relative fluorescence was measured by a spectrophotometer.

A set of control wells where no antibodies are added function as a representation of 100% infection. Then the other wells where IgY -antibodies are added are compared to the control wells and an infection ratio is calculated.

In this example, use was made of lx IgY solutions of 10 mg/ml. The parameter, relative fluorescence, measure how much fluorescent signal can be detected from the virus in each well. The lower relative fluorescence, corresponding to a lower percent infected cells, the better protection of the IgY- antibodies.

As shown in Figure 8, it is evident that RBD, ACE-2, S 1 and NRP-1 inhibit virus from infected cells.

Figure 9, A shows that a combination of IgY with specificity to NRP-1 and ACE-2 is more effective at inhibiting virus from infecting cells, when compared to treatment with IgY with specificity to either NRP-1 or ACE-2 alone. The actual effect realised is likely greater than that visualised within the corresponding graph because manner in which the antibodies in the NRP-1 and ACE-2 treatment are diluted leads to it containing half the amount of each of NRP-1 IgY and ACE-2 IgY when compared to the single treatments. Figure 10, A shows data from Figure 9, A with a specific focus on the 12 pg/ml concentration and with a comparison with an untreated control.

Figure 9, B shows that a combination of IgY with specificity to NRP-1 and RBD is more effective at inhibiting virus from infecting cells, when compared to treatment with IgY with specificity to NRP-1 alone. Figure 10, B shows data from Figure 9, B with a specific focus on the 12 pg/ml concentration and with a comparison with an untreated control.

Figure 9, C shows that a combination of IgY with specificity to ACE-2 and RBD is more effective at inhibiting virus from infecting cells, when compared to treatment with IgY with specificity to ACE-2 alone. Figure 10, C shows data from Figure 9, C with a specific focus on the 12 pg/ml concentration and with a comparison with an untreated control.

Example 9

This example shows the in vivo benefits of the treatments described herein. In particular, comparing the treatment with a composition including IgY with specificity to RBD and the “Kombo” treatment.

Treatment regime

Syrian Golden Hamsters were used as the model organism for the in vivo studies, as it is a well- established model for corona virus infections. Additionally, there is known to be a good cross-reactivity between human and hamster ACE-2 and human and hamster NRP-1 (Suresh et al., 2021, Front Pharmacol; 11:579330. doi: 10.3389/fphar.2020.579330. eCollection 2020, Ferren et al., 2021, Nature Communications, 12: 5809). Accordingly, the hamster model used herein is very applicable to the treatments described herein.

Male Syrian Golden Hamsters, age 8-10 weeks, were randomly distributed into three groups with six animals per group. At D-l (12 hrs before inoculation with virus) animals were treated as following:

• Kombo-IgY injected intraperitoneally (I.P.) with 20 mg IgY (2 ml, 10 mg/ml)

• RBD-IgY injected I.P. with 20 mg IgY (2 ml, 10 mg/ml)

• Challenge control group: not treated

At DO, 12 hrs post injection of IgY, all three groups were inoculated with 10 ³ TCID50 SARS-CoV-2 (strain: BetaCoV/Munich/BavPatl/2020) via the intranasal route. Then, 4 hours later animals were again injected with either Kombo-IgY or RBD-IgY using the same dosing as at D-l. Challenge control group were not given any treatment. At D4 all animals were euthanized and analysis on lung tissue were conducted as described in the Histopathology section.

Histopathology Histopathological analysis from selected tissues (lung and nasal turbinates) was performed for all animals. After fixation with 10% formalin, sections from left lung and left nasal turbinate were embedded in paraffin and the tissue sections were stained by H&E for histological examination. Parameters studies were alveolitis severity, bronchitis/bronchiolitis severity, rhinitis severity, tracheitis severity: 0 = no inflammatory cells, 1 = few inflammatory cells, 2 = moderate number of inflammatory cells, 3 = many inflammatory cells, alveolitis extent, 0 = 0%, 1 = <25%, 2 = 25-50%, 3 = >50%, alveolar oedema presence, alveolar haemorrhage presence, type II pneumocyte hyperplasia presence, 0 = no, 1 = yes and extent of peribronchial/perivascular cuffing, 0 = none, 1 = 1-2 cells thick, 2 = 3-10 cells thick, 3 = >10 cells thick. Results and conclusions

Table 1. Histopathological qualitative analysis of the lungs Day 4 post infection with SARS-CoV-2. The table show mean values for scoring of damages on the lungs after I.P administration of IgY twice at Day 0. Lower score indicates less damage to lung tissues. Analysis of several HE-staining show that Kombo-IgY better protects lungs compared to RBD-IgY. See the section Histopathology above for information on how the scoring is done.

Table 2. Statistical analysis of mean values from table 1; Two-tailed students t-test (p<0,05) against inoculated untreated control group. Kombo-IgY results are for several staining’s (marked in bold font) significant compared to RBD-IgY. These data show that for many indicators of lung damage in response to an infection with a coronavirus, treatment with RBD demonstrated an improvement over the negative control, with a “Kombo” treatment showing a further statistically significant improvement.

Example 10 This example shows further the in vivo benefits of the treatments described herein.

Male hamsters with age 6-8 weeks were randomly distributed into four groups with six animals per group. At DO (1 hr before inoculation with virus) animals were treated as following: • Kombo-IgY administered intranasally (LN.) with 4,8 mg IgY (70 pl, 69,2 mg/ml)

• RBD-IgY administered I.N. with 4,6 mg IgY (70 pl, 65,7 mg/ml)

• CTL infected: not treated

• CTL Non Infected: not treated

At DO, 1 hr post I.N. administration of IgY, three groups were inoculated with an intranasal dose of IO ⁵ pfu TCID50 SARS-CoV-2 (Slovakia/SK-BMC5/2020, GISAID EPI ISL 4I7879). The group CTL Non Infected was not inoculated.

Then, on DO, the groups treated with IgY were given LN. doses +1 hr and +6 hr post inoculation (equal doses as -1 hr). From DI to D3 the groups treated with IgY were given LN. doses twice a day 8 hrs apart. Body weight was registered once a day throughout the experiment. At D4 no treatment was given, and the animals were euthanized. Lung tissue from D4 were analyzed, according to the protocols described below, for total viral load (qPCR), amounts of infectious particles (TCID50) and cytokine profde was registered (qPCR).

Virus load determination in lungs by genomic qRT-PCR (Day 4)

• Quantification of viral load by RT-qPCR was done from lung using viral ORF lab gene.

• Extraction of viral RNA was performed using the Macherey Nagel NucleoSpin 96 RNA, 96- well kit for RNA purification (ref.740709.4). RNA was frozen at -80°C until qRT-PCR.

• RT was performed with the High Capacity cDNA Reverse Transcription Kit from Applied Biosystem (ref # 4368813).

• cDNA quantification by quantitative PCR was performed with primers conditions targeting ORF lab gene. Amplifications will be performed using a QuantStudio 7 Flex from Applied Biosystem and adjoining software.

• Of a note, SYBR Green technology is used (binding the fluorescent dye to double-stranded deoxyribonucleic acid (dsDNA).

Quantification of infectious particles (TCID50)

The tissue culture infective dose that causes 50% cytotoxicity (TCID50) assay is a quantitative method for assessing the infectivity of a virus stock. One TCID50 is defined as the amount of pathogen that causes death of 50% of cells (Reed and Muench, 1938), so TCID50 depends on the ability of the virus to kill the cells in culture. Infectivity is expressed as TCID50/mL/48h based on the Spearman-Karber formula. • Vero E6/TMPRSS2 cells will be counted and their viability was assessed by 0.25% trypan blue exclusion assay by ViCell apparatus.

• Cells was plated in a 96-well plate at the density of 2xl0 ⁴ cells per well in a volume of 200 pL of complete growth medium (DMEM 10% FCS).

• Cells was infected with serial dilutions of the lung homogenate (triplicate) for Ih at 37°C. Fresh medium was added for 48 hours

• 48 hours after cell infection, a MTS/PMS assay was performed (Promega ref#G5430). After discarding all supernatant, lOOpL of fresh medium and 20pL of MTS/PMS reagent was added to the culture wells. After a maximum of 4 hours, plates were read using an Elisa Plate reader and data recorded (OD value in negative cell control > 1.500).

Cytokine profiling in lungs by qRT-PCR (Day 4)

Cytokine gene expression in lungs was determined for 8 target genes (TNFa IFNy IL-2 IL-4 IL-6 IL- 10 IL-12p40 IL-21) using designated primers.

Results and conclusions

Figure 11 - Study design and treatment regime. This figure shows the design of this in vivo experiment, in which IgY was administered intranasally (IN) to a hamster, three times at day 0 (dO), and then twice a day on days 1-3 (dl-d3).

Figure 12 - Body weight. This figure shows that both “Kombo-IgY” and “RBD-IgY” prevent wight loss, with the “Kombo-IgY” performing better than the “RBD-IgY”. In the context of this experiment, “CTL non infected” was not inoculated, treated, or administered with anything.

Figure 13 - Total virus load. This figure shows the relative expression of a viral protein compared to the control of g-Actin as measured by qPCR. Both “Kombo-IgY” and “RBD-IgY” cause a statistically significant reduction of over 60% in the expression of viral protein. (CT: Cycle Threshold). The data in Figure 13 is also summarised in Table i, below. Table i. Amount of total genomic viral material, relative to a house-keeping -gene, in the lungs at Day 4.

Figure 14 - TCID50 (infectious particles at Day 4). This figure shows the amount of infectious particles in the lungs at Day 4 between the four groups. Even though total viral load (figure 13) was reduced significantly for both Kombo-IgY and RBD-IgY the total amount of infectious particles was not significantly reduced. The result indicate that a reduction of >60% of viral load is not enough to eradicate presence of living infectious viral particles. However, the cytokine profile (Figure 15) show that a lower total viral burden, as shown in Figure 13, results in a milder immune response; the animals are less affected by the SARS-CoV-2 infection. Top panel shows the histogram with a log scale at the y-axis and the lower panel with a linear y-axis.

Figure 15 - Cytokine profiling Cytokines are molecules that modulate and govern the immune response. Some cytokines upregulate the immune response while others downregulate it. Thus, there is a very complex mechanism regulating the intensity of the immune response to a given pathogen. For COVID- 19 in humans it is observed that IL6 is a biomarker for the severity of the illness, as explained below. An upregulated IL6 response means that the patient’s immune response is reacting strong to the infection. IL6 alone could not cause a cytokine storm but when looking at the profile of cytokine expression it can be used to evaluate the severity of COVID- 19 and whether a patient needs intensive care.

It is known that IL-6 is a marker for a severe coronavirus infection (Santa Cruz et al., 2021, Front Immunol; 12:613422; Broman et al., 202, Ann Med;53(l):410-412; Sabaka et al., 2021, BMC Infectious Diseases, 21, 308; Tang et al., 2021, Medicine: 100, 22: p e26131). Figure 15 shows that treatment with RBD leads to a reduction in IL-6 with the Kombo treatment leading to a further reduction in IL-6, indicating that the treatments described herein have an impact on severe coronavirus infections. The invention is further described in the following numbered paragraphs:

1. IgY antibody composition, wherein the IgY antibody composition is obtained by immunization of an egg-laying poultry (layer) with one or more antigens selected from Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-l) antigen, SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, and SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

2. IgY antibody composition in accordance with paragraph 1, wherein the IgY antibody composition is obtained by immunization of an egg-laying poultry (layer) with a combination of two or more antigens selected from Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-l) antigen, SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen and SARS- CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, preferably a combination of Angiotensin Converting Enzyme 2 (ACE-2) antigen and one or more antigens selected from Neuropilin 1 (NRP-l) antigen, SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen and SARS- CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, more preferably a combination of Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-l) antigen and optionally one or more antigens selected from SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen and SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen.

3. IgY antibody composition for use in prevention and/or treatment of an infection of a coronavirus in an animal, wherein the IgY antibody composition is obtained by immunization of an egg-laying poultry with one or more antigens selected from Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-l) antigen, SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, and SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

4. IgY antibody composition for use in prevention and/or treatment of an infection of a coronavirus in an animal in accordance with paragraph 3, wherein the IgY antibody composition is obtained by immunization of an egg-laying poultry with two of more antigens selected from Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-l) antigen, SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, and SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen, preferably a combination of Angiotensin Converting Enzyme 2 (ACE-2) antigen and one or more antigens selected from Neuropilin 1 (NRP-l) antigen, SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen and SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, more preferably a combination of Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-l) antigen and optionally one or more antigens selected from SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen and SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen.

5. IgY antibody composition in accordance with paragraph 1 or 2, or IgY antibody composition for use in prevention and/or treatment of an infection of a coronavirus in an animal in accordance with paragraph 3 or 4, wherein the IgY antibody composition is obtained by immunization of an egg-laying poultry (layer) with Angiotensin Converting Enzyme 2 (ACE- 2) antigen.

6. IgY antibody composition in accordance with paragraph 1 or 2, or IgY antibody composition for use in prevention and/or treatment of an infection of a coronavirus in an animal in accordance with paragraph 3 or 4, wherein the IgY antibody composition is obtained by immunization of an egg-laying poultry with Neuropilin 1 (NRP-l) antigen.

7. IgY antibody composition in accordance with paragraph 1 or 2, or IgY antibody composition for use in prevention and/or treatment of an infection of a coronavirus in an animal in accordance with paragraph 3 or 4, wherein the IgY antibody composition is obtained by immunization of an egg-laying poultry with SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen.

8. IgY antibody composition in accordance with paragraph 1 or 2, or IgY antibody composition for use in prevention and/or treatment of an infection of a coronavirus in an animal in accordance with paragraph 3 or 4, wherein the IgY antibody composition is obtained by immunization of an egg-laying poultry with SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

9. A pharmaceutical composition comprising an IgY antibody composition according to any of the paragraphs 1-8, and at least one medical compatible adjuvant.

10. Egg yolk from an egg from an egg-laying poultry (layer) for use in prevention and/or treatment of an infection of a coronavirus in an animal, wherein the egg yolk comprises IgY antibodies obtained by immunization of the egg-laying poultry with one or more antigens selected from Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-l) antigen, SARS- CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, and SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen. 11. Egg yolk from an egg from an egg-laying poultry (layer) for use in prevention and/or treatment of an infection of a coronavirus in an animal in accordance with paragraph 10, wherein the egg yolk comprises IgY antibodies obtained by immunization of the egg-laying poultry with two or more antigens selected from Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-l) antigen, SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, and SARS- CoV-2 Spike Glycoprotein 1 (S 1) antigen, preferably a combination of Angiotensin Converting Enzyme 2 (ACE-2) antigen and one or more antigens selected from Neuropilin 1 (NRP-l) antigen, SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen and SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, more preferably a combination of Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-l) antigen and optionally one or more antigens selected from SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen and SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen.

12. A method of in vitro diagnosis, wherein an IgY antibody composition in accordance with any of paragraphs 1-8 is contacted with a biological sample from an animal, and wherein a neutralization or complexation of the IgY antibodies with a virus in the sample indicates that the sample contains corona virus or other RNA viruses.

13. The IgY antibody composition for use in prevention and/or treatment of an infection of a coronavirus in an animal in accordance with any one of paragraphs 3 to 8, or the egg yolk from an egg from an egg-laying poultry (layer) for use in prevention and/or treatment of an infection of a coronavirus in an animal in accordance with any one of paragraph 10 and 11, or the method according to paragraph 12, wherein the infection and/or virus is caused by SARS-CoV-2.

14. The IgY antibody composition for use in prevention and/or treatment of an infection of a coronavirus in an animal in accordance with any one of paragraphs 3 to 8 and 13, or the pharmaceutical composition comprising an IgY antibody composition according to paragraph

9, or the egg yolk from an egg from an egg-laying poultry (layer) for use in prevention and/or treatment of an infection of a coronavirus in an animal in accordance with any one of paragraph

10, 11 and 13, or the method in accordance with any one of paragraph 12 and 13, wherein the IgY antibody composition is administered to an animal as an injection, nose spray, inhalation or an aerosol (nebulization). The IgY antibody composition for use in prevention and/or treatment of an infection of a coronavirus in an animal in accordance with any one of paragraphs 3 to 8 and 13 to 14, or the pharmaceutical composition comprising an IgY antibody composition according to paragraph 9 and 14, or the egg yolk from an egg from an egg-laying poultry (layer) for use in prevention and/or treatment of an infection of a coronavirus in an animal in accordance with any one of paragraphs 10, 11 and 13 to 14, or the method in accordance with any one of paragraph 12 to 14, wherein the animal is a human. The IgY antibody composition in accordance with any one of paragraphs 1, 2 and 5 to 8, or the IgY antibody composition for use in prevention and/or treatment of an infection of a coronavirus in an animal in accordance with any one of paragraphs 3 to 8 and 13 to 15, or the pharmaceutical composition comprising an IgY antibody composition according to any one of paragraphs 9, 14 and 15, or the egg yolk from an egg from an egg-laying poultry (layer) for use in prevention and/or treatment of an infection of a coronavirus in an animal in accordance with any one of paragraphs 10, 11 and 13 to 15, or the method in accordance with any one of paragraphs 12 to 15, wherein the IgY antibody composition and/or IgY antibodies comprises polyclonal antibodies. The IgY antibody composition in accordance with any one of paragraphs 1, 2, 5 to 8 and 15 to

16, or the IgY antibody composition for use in prevention and/or treatment of an infection of a coronavirus in an animal in accordance with any one of paragraphs 3 to 8 and 13 to 16, or the egg yolk from an egg from an egg-laying poultry (layer) for use in prevention and/or treatment of an infection of a coronavirus in an animal in accordance with any one of paragraphs 10, 11, and 13 to 16, wherein the immunization of the egg-laying poultry takes place by administering one or more antigens by one or more injections. The IgY antibody composition in accordance with any one of paragraphs 1, 2, 5 to 8 and 15 to

17, or the IgY antibody composition for use in prevention and/or treatment of an infection of a coronavirus in an animal in accordance with any one of paragraphs 3 to 8 and 13 to 17, or the egg yolk from an egg from an egg-laying poultry (layer) for use in prevention and/or treatment of an infection of a coronavirus in an animal in accordance with any one of paragraphs 10, 11, and 13 to 17, wherein the immunization of the egg-laying poultry further comprises collecting egg of the egg-laying poultry, obtaining egg yolk from the egg, extracting and isolating the IgY antibody composition and/or IgY antibodies from the egg yolk and purifying the IgY antibody composition and/or IgY antibodies. The invention is further described in the following letered paragraphs:

A. IgY antibody composition, wherein the IgY antibody composition is obtained by immunization of an egg-lying poultry (layer) with an Angiotensin Converting Enzyme 2 (ACE-2) antigen.

B. IgY antibody composition in accordance with paragraph A, wherein the IgY antibody composition is obtained by immunization of an egg-lying poultry (layer) with a combination of Angiotensin Converting Enzyme 2 (ACE-2) antigen and SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen.

C. IgY antibody composition in accordance with paragraph A, wherein the IgY antibody composition is obtained by immunization of an egg-lying poultry (layer) with a combination of Angiotensin Converting Enzyme 2 (ACE-2) antigen and SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

D. IgY antibody composition in accordance with paragraph A, wherein the IgY antibody composition is obtained by immunization of an egg-lying poultry (layer) with a combination of Angiotensin Converting Enzyme 2 (ACE-2) antigen and Neuropilin 1 (NRP-1) antigens.

E. IgY antibody composition in accordance with paragraph A, wherein the IgY antibody composition is obtained by immunization of an egg-lying poultry (layer) with a combination of Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-1) antigen and SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

F. IgY antibody composition in accordance with paragraph A, wherein the IgY antibody composition is obtained by immunization of an egg-lying poultry (layer) with a combination of Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-1) antigen and SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen.

G. IgY antibody composition in accordance with paragraph A, wherein the IgY antibody composition is obtained by immunization of an egg-lying poultry (layer) with a combination of Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP-1) antigen, SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen and SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen.

H. IgY antibody composition, wherein the IgY antibody composition is obtained by immunization of an egg -lying poultry (layer) with Neuropilin 1 (NRP-1) antigens.

I. IgY antibody composition in accordance with paragraph H, wherein the IgY antibody composition is obtained by immunization of an egg-lying poultry (layer) with Angiotensin Converting Enzyme 2 (ACE-2) antigens and Neuropilin 1 (NRP-1) antigens.

J. IgY antibody compositions in accordance with paragraph H, wherein the IgY antibody composition is obtained by immunization of an egg-lying poultry (layer) with a combination of two or more antigens selected from Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP- 1) antigen, SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen and SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen.

K. IgY antibody composition, wherein the IgY antibody composition is obtained by immunization of an egg-lying poultry (layer) with SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen.

L. IgY antibody composition in accordance with paragraph K, wherein the IgY antibody composition is obtained by immunization of an egg-lying poultry (layer) with SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigens and SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

M. IgY antibody compositions in accordance with paragraph K, wherein the IgY antibody composition is obtained by immunization of an egg-lying poultry (layer) with a combination of two or more antigens selected from Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP- 1) antigen, SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen and SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen.

N. IgY antibody composition in accordance with paragraph K, wherein the IgY antibody composition is obtained by immunization of an egg-lying poultry (layer) with SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

O. IgY antibody composition, wherein the IgY antibody composition is obtained by immunization of an egg-lying poultry (layer) with SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigens and SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

P. IgY antibody compositions in accordance with paragraph K, wherein the IgY antibody composition is obtained by immunization of an egg-lying poultry (layer) with a combination of two or more antigens selected from Angiotensin Converting Enzyme 2 (ACE-2) antigen, Neuropilin 1 (NRP- I) antigen, SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen and SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen.

Q. IgY antibody composition in accordance with paragraphs A, H, K or N, wherein the composition comprises several different antibodies which will bind to different epitopes of the corona virus.

R. IgY antibody composition in accordance with paragraphs A, H, K or N, wherein the IgY antibodies are polyclonal.

S. IgY antibody composition in accordance with paragraphs A, H, K or N, wherein the immunization of the egg-laying poultry is conducted by providing the combination of antigens as an aerosol.

T. IgY antibody composition in accordance with paragraphs A, H, K or N, wherein the immunization of the egg-laying poultry is conducted by providing the combination of antigens as an injection. U. IgY antibody composition in accordance with paragraphs A, H, K or N, wherein the immunization of the egg-laying poultry is conducted by providing the combination of antigens by an intramuscular injection.

V. IgY antibody composition according to paragraphs A, H, K or N, wherein the antigens specific for corona viruses is a cocktail of antigens specific for corona 3 viruses.

W. IgY antibody composition in accordance with paragraphs A, H, K or N, wherein the Angiotensin Converting Enzyme 2 (ACE-2) antigen is Angiotensin Converting Enzyme 2 expressed in HEK293 cells or the ACE-2 protein, available as Cat. 90211-C02H from SinoBiological.

X. IgY antibody composition in accordance with paragraphs A, H, K or N, wherein the Neuropilin 1 (NRP-l) antigens is the sequence Phe 22 - Lys 644 expressed in HEK293, available from 2bScientific.

Y. IgY antibody composition in accordance with paragraphs A, H, K or N, wherein the Spike antigens are the SARS-CoV-2 Spike Glycoprotein (SI (seql-674)) SHFc-Tag (HEK293) or SARS- CoV-2 Spike Glycoprotein (S2 (seq685-1211)) SHFc-Tag (HEK293), or a combination of both.

Z. IgY antibody composition for the use in prevention and/or treatment of an infection of a coronavirus in an animal, wherein the IgY antibody composition is obtained by immunization of an egg-laying poultry with an Angiotensin Converting Enzyme 2 (ACE-2) antigen.

AA. IgY antibody composition for the use in prevention and/or treatment of an infection of a coronavirus in an animal, wherein the IgY antibody composition is obtained by immunization of an egg-laying poultry with Neuropilin 1 (NRP-l) antigens.

BB. IgY antibody composition for the use in prevention and/or treatment of an infection of a coronavirus in an animal, wherein the IgY antibody composition is obtained by immunization of an egg-laying poultry with SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen.

CC. IgY antibody composition for the use in prevention and/or treatment of an infection of a coronavirus in an animal, wherein the IgY antibody composition is obtained by immunization of an egg-laying poultry with SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen.

DD. IgY composition in accordance with any of the paragraphs Z-CC, wherein the IgY antibody composition is obtained by immunization of an egg-laying poultry with a combination of two or more of Angiotensin Converting Enzyme 2 (ACE-2) antigen, SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen, SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen and Neuropilin 1 (NRP-l) antigens.

EE. IgY antibody composition in accordance with any of the paragraphs Z- CC, wherein said animal is a human.

FF. IgY antibody composition in accordance with any of the paragraphs Z- CC, wherein the disease caused by the corona virus is COVID- 19. GG. IgY antibody composition in accordance with any of the paragraphs Z-CC, wherein the composition comprises several different antibodies which will binds to different epitopes of the corona virus.

HH. IgY antibody composition in accordance with any of the paragraphs Z-CC, wherein the IgY antibodies are polyclonal.

II. IgY antibody composition in accordance with any of the paragraphs Z-CC, wherein the immunization of the egg-laying poultry is conducted by providing the combination of antigens as an injection.

JJ. IgY antibody composition in accordance with any of the paragraphs Z-CC, wherein the immunization of the egg-laying poultry is conducted by providing the combination of antigens by an intramuscular injection.

KK. IgY antibody composition according to any of the paragraphs Z-CC, wherein the antigens specific for corona viruses is a cocktail of antigens specific for corona 3 viruses.

LL. A pharmaceutical composition comprising an IgY antibody composition according to any of the paragraphs A-KK, and at least one medical compatible adjuvant.

MM. Egg yolk from an egg from an egg-laying poultry (layer) for use in prevention and/or treatment of an infection of a coronavirus in an animal wherein the egg yolk comprises IgY antibodies obtained by immunization of the egg-laying poultry with an Angiotensin Converting Enzyme 2 (ACE-2) antigen. NN. Egg yolk from an egg from an egg-laying poultry (layer) for use in prevention and/or treatment of an infection of a coronavirus in an animal wherein the egg yolk comprises IgY antibodies obtained by immunization of the egg-laying poultry with a SARS-CoV-2 Spike Glycoprotein 1 (SI) antigen 00. Egg yolk from an egg from an egg-laying poultry (layer) for use in prevention and/or treatment of an infection of a coronavirus in an animal wherein the egg yolk comprises IgY antibodies obtained by immunization of the egg-laying poultry with Neuropilin 1 (NRP-l) antigens.

PP. Egg yolk from an egg from an egg-laying poultry (layer) for use in prevention and/or treatment of an infection of a coronavirus in an animal wherein the egg yolk comprises IgY antibodies obtained by immunization of the egg-laying poultry with SARS-CoV-2 Spike 1 Receptor Binding Domain (RBD) antigen.

QQ. A method of in vitro diagnosis, wherein an IgY in accordance with any of the paragraphs A-PP are contacted with a biological sample from an animal, and wherein a neutralization or complexation of the IgY with a virus in the sample indicates that the sample contains corona virus or other RNA viruses.

RR. A method according to paragraph QQ, wherein the virus is SARS-COV-2.