CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. provisional patent application no.

62/187,501, filed July 1, 2015, the disclosure of which is incorporated herein by reference.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

[0002] This invention was made with government support under Contract No.

N00014-13-1-0595, awarded by the Office of Naval Research and under Contract No.

W911NF-14-P-0012, awarded by the U.S. Army/ARO. The Government has certain rights in the invention.

FIELD

[0003] The present disclosure relates generally to devices and methods for collecting and/or processing of biological samples. BACKGROUND

[0004] Current sample collection devices (SCD's) for cellular materials, especially those destined for forensic analysis, have not kept pace with the trace quantity (i.e., nanogram to -100 pico-gram for nucleic acids) required for sensitive and reliable genetic profiling technology. The SCD's swab fiber materials are typically manufactured from the classic cotton, to more recently with rayon, nylon, foam and inorganic polysilane. These generally perform relatively well in the collection step but, to varying degrees, typically lose 50-80% of in the eventual release and retrieval of the trace material. The inherent problem lies in the fiber material of the SCD's as they trap and bind irreversibly to biological material. Thus, there is an ongoing and unmet need for improved compositions, devices and methods for collecting and processing biological samples. The present disclosure addresses this need.

SUMMARY

[0005] In general the present disclosure relates to sample collection devices (SCDs) comprising a protein component. The protein component is suitable for separating biological macromolecules from a test location. Generally, the protein component forms a complex comprising the protein component and the biological macromolecule(s), wherein the protein component and the biological macromolecule(s) are non-covalently associated with one another. Biological macromolecules that can be separated from a test location and tested according to the present disclosure include but are not necessarily limited to nucleic acids (i.e. DNA and RNA polynucleotides), proteins, polypeptides, peptides, carbohydrates, lipids, and combinations thereof.

[0006] In certain implementations a protein component of the sample collection device comprises Squid Ring Teeth (SRT) protein. The SRT protein may be provided in a variety of forms, including but not necessarily as a foam or a fiber. The protein component may be provided such that it is the only part of the SCD that forms a complex with the macromolecules, but the SRT may be in physical association with other components of the SCD, such as a wooden or plastic handle or other non-protein components, and may be in physical associate with a non-protein substrate, such as cotton, nylon, rayon, etc. In certain aspects the protein component is an SRT protein that is in the form of a foam, or as fibers. The foam or fibers or other configurations of the SRT protein may comprise or consist essentially of or consist of SRT.

[0007] The SCD can be provided in various arrangements, such as a pestle coating, the pestle being constructed of any solid material including but not necessarily limited to plastics, metals and wood. The SCD can be configured such that it and/or its SRT protein- containing composition is present in a flexible material, such as an individual swipe or swab, or an adhesive material, such as a tape. The SCDs can be provided as individual members, or in sheets from which separate SCDs can be removed and used in methods of the disclosure. In non-limiting implementations, the SCD can comprise a filter device, and/or a spin column device. In non-limiting examples the SRT-protein can be provided as a plurality of particles, such as beads, microparticles, nanoparticles, etc., or in a matrix. The disclosure includes SCDs that comprise biological macromolecules separated from a test location, wherein the biological macromolecules may be in a complex with the protein component of the device. But in certain embodiments, the macromolecules may be comprised by cells, and the device can comprise the cells comprising the macromolecules.

[0008] In another aspect the disclosure includes kits that contain an SCD as described herein. The kits can comprise reagents, such as liquid buffers, used in methods of the disclosure.

[0009] In another aspect the disclosure provides a method for analyzing a test sample for the presence or absence of biological macromolecules, the method comprising contacting a test location with a SCD of this disclosure, and removing the sample collection device from the location such that the macromolecules if present are separated from the location. The test sample can then be processed to determine the presence or absence of the biological macromolecule using a variety of approaches that are well known in the art, including but not limited to determining the sequences of polynucleotides and/or the amino acid content of and/or the identity of proteins. Thus, the biological macromolecules from the test sample can be characterized. BRIEF DESCRIPTION OF THE FIGURES

[0010] Figure 1. (a) Liquid absorbance of SRT foam, (b) XRD data shows semi- crystalline architecture of the SRT, (c) Electron microscopy shows the porous architecture of the SRT foam.

[0011] Figure 2. SRT binding DNA in (a) denaturing and (b) non-denaturing binding conditions are shown. Triggered mock experiment to demonstrate complete dissolution of SRT matrix in G-HC1 and phenol and other commercially available swab type SCD's (i.e, Cotton, Rayon, Puritan, Nylon, Polyester, Silk) in (c) guadiamine-HCl and (d) Phenol.

[0012] Figure 3. Photographic depictions of swab preparation via electrospinning and dip-coatings, (a) SRT -fiber mat. (b) Sample collection device with wood scraper (i.e., a handle), (c) SRT+ Cotton fibers, (d) Blood + SRT coated cotton.

DETAILED DESCRIPTION

[0013] Unless defined otherwise herein, all technical and scientific terms used in this disclosure have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains.

[0014] Every numerical range given throughout this specification includes its upper and lower values, as well as every narrower numerical range that falls within it, as if such narrower numerical ranges were all expressly written herein.

[0015] The present disclosure provides improved biological collection materials, devices and methods of using the same. All compositions of matter, composites, devices, processes, and results disclosed herein are encompassed by the instant disclosure.

[0016] In certain embodiments, the disclosure encompasses a device comprising at least one organic polymer, i.e., at least one polypeptide, wherein the device is i) suitable for separating compositions of matter from a test location to obtain a test sample, wherein the test sample comprises or is suspected of comprising a composition of matter, and ii) processing the test sample. In embodiments, the compositions of matter in the test samples comprise biological macromolecules. In embodiments, processing the test sample comprises: a) determining the presence or absence of the biological macromolecule, and/or b) determining at least one physical characteristic of the biological macromolecule. Determining

characteristics of macromolecules can comprise characterization of cells, including but not necessarily limited to prokaryotic and eukaryotic cells, and can also include characterizing viruses.

[0017] Biological macromolecules include but are not necessarily limited to nucleic acids, proteins, polypeptides, peptides, carbohydrates, lipids, and combinations thereof. In embodiments, the biological macromolecules comprise compounds that have biological activity, and thus include but are not necessarily limited to drug compounds and/or other small molecules that may have pharmacokinetic or other biological affects in, for example, a mammal, through any of a wide variety of processes.

[0018] In certain embodiments, the disclosure comprises contacting a test location with a sample collection device (SCD) as described herein to obtain a combination of the device and a test sample, and performing at least one process step to characterize a composition of matter in the test sample. Thus, in embodiments, the disclosure includes a sample collection device as described herein, wherein the protein constituent of the device is a component of a complex that comprises the protein and a plurality of non-covalently bound biological macromolecules. Any composition of matter formed during a processing step described herein is also encompassed by this disclosure.

[0019] In general, at least one processing step comprises separating the composition of matter from the SCD. Separating the composition of matter comprises any suitable process and/or means for dissociating the composition of matter from the device so that the composition of matter can be analyzed. In certain aspects, separating the composition of matter from the device comprises diffusion, washing with a solvent (i.e., elution), affinity separation, size separation, chromatography, centrifugation, electrophoresis, or any by using any other techniques and/or reagents that will be apparent to those skilled in the art given the benefit of the present disclosure. In one embodiment, the device itself is disrupted, disintegrated or dissolved, such as in a suitable buffer, and the composition of matter collected by the device is separated from the device component(s). In embodiments, the composition of matter can be isolated and/or purified to any desired degree of purity.

[0020] In a non-limiting illustration to demonstrate one of many possible approaches, a device comprising a test sample according to this disclosure comprises nucleic acids. In one embodiment the nucleic acid is DNA. The device comprising DNA can be processed to analyze the DNA, such as by determining the sequence of the DNA. The sequence of the DNA in certain embodiments can be determined while the DNA is still present in the device/test sample combination, or the DNA can be separated from the device using any suitable approach, various illustrative examples of which are described further below. In embodiments, the sequence of the DNA is determined to support an inference or conclusion about the source of the sample. In non-limiting embodiments the disclosure is thus suitable for a wide array of analytic applications, including but not necessarily limited to analysis in conjunction with forensics, such as determining trace amounts of DNA from a crime scene, genotypic- analysis, such as to determine or predict a trait in an organism, population genetics, human and veterinary medicine, such as for diagnosing or aiding in a diagnosis of a disease, and/or for making a prognosis or treatment recommendation or for staging a diseases, epidemiology, pedigree and breeding analysis, agricultural applications in the animal and plant science areas, water analysis, food analysis, such as for analysis of food safety and the presence or absence of pathogens, paleontology and archeology, such as to determine trace amounts of DNA from such samples, parasitology and other infectious disease analysis, and many other applications wherein determining the presence or absence of DNA and/or its sequence would be desirable. In embodiments, the DNA or other nucleic acid in the test sample is amplified, and the amplified nucleic acids, i.e., amplicons, are analyzed. In embodiments, nucleic acid analysis comprises determining a polynucleotide sequence, and can comprise determining all of the contiguous nucleotides in a

polynucleotide, or a subset of polynucleotides, a repeated sequence, an insertion, a deletion, or as few as one nucleotide, such as in determining the presence of absence of, for example, a single nucleotide polymorphism (SNP), such as to detect a marker, an allele, a haplotype, or any other genetic or phenotypic indicator that is correlated with the SNP. Any suitable approach for determining polynucleotide sequences can be used, and many are known in the art and are commercially available. In embodiments, the polynucleotide analyzed according to the instant disclosure comprises RNA, including but not limited to mRNA, miRNA, and snoRNA.

[0021] As discussed above, in certain aspects, the disclosure includes contacting a test location with a device as described herein to obtain a combination of the device and a test sample, and performing at least one process step to separate a composition of matter in the test sample from the device. In a non-limiting illustration, DNA or RNA can be separated from the device as further described below, amplified if desired, and cloned into any suitable cloning vector, many of which are known in the art and are commercially available. The disclosure also contemplates creating libraries of samples, wherein the library comprises a plurality of distinct compounds separated from SCD device/sample combinations. Thus libraries of amplified nucleic acids are included.

[0022] Those skilled in the art will recognize that any other composition of matter can be tested for in a test sample obtained according to this disclosure. In embodiments, the test sample is tested for the presence or absence of a protein that is distinct from the SRT protein that forms a component of a device of this disclosure as described further below, including but not necessarily limited to protein or peptide disease markers, allergens or other antigens, peptide hormones, receptor ligands, chemokines, cytokines, etc.

[0023] In other embodiments, the test sample is tested for the presence or absence of a carbohydrate, a lipid, a steroid, a small molecule, such as a drug, including but not necessarily limited to therapeutic drugs and drugs of abuse, cellular components, organic and inorganic components, etc.

[0024] In embodiments, devices of this disclosure are suitable for separating any biological macromolecule from any test location. In embodiments, the test location comprises a solid surface, a porous surface, a gas, vapor, or a liquid. In embodiments, the test location is present on an individual, such as a mammal.

[0025] In embodiments, the test sample comprises a biological sample, including but not limited to a sample of a tissue, cells, a biological fluid, a skin sample, etc. In

embodiments, the test sample comprises blood, plasma, serum, urine, saliva, tears, semen, sperm, cerebrospinal fluid, tissue (i.e., a biopsy), hair, hair follicles, bone, teeth, skin, nucleated cells of any kind, including but not limited to epithelial cells, etc.

[0026] In certain embodiments, a sample collection device of the present disclosure is insoluble or very poorly soluble in water having a pH range of 6-8 and is stable at room temperature (i.e., approximately 70°F) for prolonged periods, i.e., for at least at period of one month, up to and including a period of at least one year, including all days and ranges of days between one month and one year, or for a period longer than one year. In embodiments, a sample collection device of the present disclosure collects an amount of biological material that is between 50%- 100%, inclusive, and including all integers and ranges of integers there between, of the biological material present on a test surface and with which the sample collection device comes into contact. In embodiments, the amount of biological material that is subsequently released from the test device and/or can be tested using any suitable technique comprises between 50%-100%, inclusive, and including all integers there between, of the biological material initially collected by the device. In one aspect, a sample collection device as provided herein will capture cellular material with efficiency greater than 95%, and release at least 80% of the cellular material in, for example, less than one hour. [0027] In general, the collection devices of this disclosure comprise Squid Ring

Teeth Protein (SRT). In one non-limiting embodiment discussed further below, squid ring teeth (SRT) is a structural protein extracted from tentacles of squid suction cups, which exhibits an unusual and reversible transition (i.e., thermoplastic) from a solid to a rubber, and can be thermally shaped into any 3D geometry (e.g. foams, fibers, colloids, and thin films). SRT thus is comprises a unique class of high-performance elastomeric proteins which have excellent mechanical properties, in wet and dry conditions that exceed most natural and synthetic polymers, while having the unique capability of being thermally remodeled into various functional forms such as films, foams, and fibers suitable for SCD's. SRT from any species of squid can be used. In certain embodiments the SRT is from Loligo vulgaris. In certain embodiments the SRT is full length SRT protein. In certain embodiments the SRT comprises or consists of an 18kDa SRT protein, or 22 kDa SRT protein, or a mixture of those proteins, which may further comprise a full length SRT protein. In certain embodiments the SRT component of an SCD of this disclosure comprises or consists of SEQ ID NO: 1, or SEQ ID NO:2. Mixtures of proteins comprising or consisting of SEQ ID NO: 1 and SEQ ID NO:2 are included.

[0028] In an embodiment, a test device of this disclosure consists entirely of SRT protein, or consists essentially of SRT protein, or comprises of SRT protein and at least one additional composition of matter, such as at least one additional organic or inorganic composition. In an embodiment, the additional composition comprises an inorganic compound. In an embodiment the additional composition is an inert material. In

embodiments, the device comprises one or more reagents to aid in detection/preservation of biological macromolecules. In embodiments, the device comprises or is provided with a deoxyribonucleases (DNases) inhibitor and/or an RNase inhibitor, or a nuclease inhibitor cocktail. In embodiments, a chelating agent is included. In embodiments, one or more reagents to aid in, for example, nucleic acid separation from cells can be included, such as guanidinium thiocyanate (GITC), or detergents, or protein denaturing agents.

[0029] SCDs of the present disclosure can be provided in a wide variety of physical confirmations. In certain, non-limiting embodiments, the SCD is provided as a foam. In other embodiments, the SCD comprises SRT protein as a component of a matrix with at least one other material. In embodiments, the SCD is provided as a porous solid matrix suitable for layering onto a member for holding the sample collection material. In embodiments, a SRT protein-comprising composition of this disclosure is layered onto an article for positioning and using the composition to contact a test surface. In embodiments, the SCD comprises a pestle. In embodiments, a SRT protein-containing composition of this disclosure is provided as a fiber, such as to coat a polypropylene pestle, or pestle of any solid material including but not necessarily limited to metal and wood. In an embodiment, the SRT protein- containing composition is present in a flexible material, such as an individual swipe, or an adhesive material, such as a tape. The SCDs can be provided as individual members, or in sheets containing a plurality of members. In embodiments, the SCD can comprise a filter device, and/or a spin column device. In embodiments, the SRT -protein comprising composition is a plurality of particles, such as beads, microparticles, nanoparticles, and the like. In embodiments, the SCD comprises fibers, such as a matrix of fibers comprising or consisting of a recombinant or native SRT protein or protein complexes.

[0030] In embodiments the disclosure includes kits that comprise any SCD described herein. The kits can further comprise any one or combination of reagents suitable for sample collection, and/or separation from the SCD, and/or for biological sample preservation and/or analysis. In embodiments, an SCD of this disclosure is provided with, for example, a suitable collection and/or extraction buffer or other solution. In the kit, the SCD and other reagents can be provided in separate, sealed, sterile containers. In embodiments, the kits comprise a sample collection device adapted for use in DNA analysis. In embodiments, the kits comprise a single SCD, or more than one SCD. In embodiments, the kits comprise a composition comprising or consisting of a protein that is suitable for assembling into an SCD, such as a powder. The kits can comprise instructions, such as printed material, for sample collection and/or preservation and/or transportation and/or processing, or for assembling the SCD from a pre-SCD preparation, such as a powder, gel, or other formable composition, and/or for storage of the SCD with or without a test sample as a component of it. In embodiments the disclosure includes an article of manufacture comprising packaging material comprising a kit of this disclosure. The article of manufacture can comprise printed material, such as a label or printed paper that describes the contents of the article.

[0031] In embodiments, the present disclosure comprises analyzing a test sample using an SCD as described herein to obtain a characterization of a biological macromolecule in the sample, and fixing the characterization in a tangible medium, such as a paper report, or a digitized file. In embodiments, the method further comprises communicating the characterization to a recipient, such as a law enforcement official, a health care provider, or any other individual with an interest in the characterization.

[0032] In embodiments, a device of the present disclosure comprises recombinant

SRT which as described above is a structural protein extracted from tentacles of squid suction cups. In embodiments, the SRT comprises a protein comprising an amino acid sequence disclosed herein, or a fragment of such an amino acid sequence suitable for use in an SCD of this disclosure. Benefits of using SRT in the current approach include but are not necessarily to: (i) reversible assembly due to physically cross-linked structure of β-sheets, (ii) strong mechanical properties (i.e., -lGPa modulus) due to semi-crystalline molecular structure, (iii) reduced weight compared to carbon fibers (density of 1.3 vs. 1.8), (iv) reduced energy demands for materials production (-20 kcal/g v.s. -10 kcal/g for plastic and recombinant expression respectively), (v) biodegradability, (vi) processing versatility (e.g. extrusion, injection molding, casting, etc.), and (vii) novel materials' synthesis strategy based on "sequence -structure-property" relationship with tunable physical properties. Based on these properties, the present disclosure provides improvements relative to previously available plastic and cotton based swabs. In particular, in current approaches, 50-80% of the collected material is lost as the material is not available for extraction, possibly due to irreversible binding and trapping in the typical sample collection device fiber material. Thus, an innovation of the present disclosure is the capability for greatly improved release and recovery of the collected biological material. In certain embodiments, these and other improvements are obtained by formation of non-covalent bonds between the captured material and the sample collection device material, and triggering the sample collection device itself to dissolve by exposure to an appropriate, controlled stimulus (e.g. a change in pH, a chemical composition, or other change) thus releasing quantitatively the bound material. Such capability is considered to be of particular value in certain aspects of the disclosure, such as for touch DNA (i.e., tape lifting) collection methods. In embodiments, such as for DNA extraction, standard protocols can be used, such as either the well-known method of phenol extraction, or the non-organic approaches using, for example, chaotropic salt, such as G-HC1. In this regard, it is demonstrated herein that SRT protein dissolves or softens in chemical solvents (e.g., 7M Guanidinium Hydrochloride (G-HC1), phenol) or by pH titration below 3.

[0033] The current approach is accordingly superior to many of the currently available SCD's swab fiber materials, which comprise cotton, rayon, nylon, foam and inorganic polysilane. These have inherent deficiency of not releasing 100% of the captured material, which become trapped and irreversibly bound to the SCD's fiber matrix. Working with trace material especially in a forensic setting requires maximum DNA capture and retrieval. In this disclosure, we demonstrate the use of SRT structural protein as strong viable alternate for construction of the SCD's fiber material that will capture biological material with high efficiency, and more significantly release the cellular material quantitatively in solution for downstream DNA extraction.

[0034] The following specific examples are provided to illustrate the invention, but are not intended to be limiting in any way.

EXAMPLES

[0035] At least two methods can be used to create SRT foams. In the first method, foams can be made by forming gas bubbles with the use of a blowing agent (e.g.,

hydrocarbons such as pentane, isopentane, cyclopentane or liquid C02). This method takes advantages of the thermoplastic properties of SRT. In the second method, a solution-casting method can be used with a salt. SRT can be dissolved in hexafloroisopropanol (HFIP) and incubated with a salt (e.g., 20: 1 ratio of NaCl:SRT) that is stable in HFIP. The solvent can be evaporated at room temperature. Remaining salt can be dissolved in water, which will produce a porous protein structure.

[0036] The foam can be shown to exhibit stimuli responsive dissolution. In this regard, SRT is a structural protein that is hold together with physical crosslinking of β-sheets. SRT can be dissolved by titrating the pH or by the choice of a polar solvent. Intermolecular forces explain the mechanism of SRT dissolution up on pH change. Compared to other bio- derived thermoplastics (e.g., keratin, resilin, elastin), the proteinaceous structure of the SRT does not have any covalent cross-linker and hence, dissolves in polar protic or weak alkaline/acidic buffers, and SRT reversibly agglomerates if the solvent is removed. The isoelectric point of the protein film is 6.7. Therefore, the net charge on the native SRT protein film is close to zero at physiological buffers (e.g., pH, ~7), which is related to the major charged residue in native SRTs being histidine whose pKa is 6.5. The net charge increases as the pH is lowered, and finally breaks the balance of β-sheets, which is hold together by hydrogen bonds. Depending on the choice of recombinant SRT protein (where the histidine amino acid content varies), the isoelectric point changes (e.g., from 7 to 9). All recombinant proteins can be tested to optimize the stimuli responsive dissolution of the foam.

[0037] It is expected that embodiments of the present disclosure will function to capture and trigger release of human cells from SCD. It is feasible to expect capture of human cells with 95% efficiency in a range of 25K to 400K cells. Saliva and whole blood sample can be diluted in 10 mM PBS (noting that whole blood with 3-7 x 106 white blood cells/mL). Varying dilutions (1 : 10, 1 :50 and 1 : 100) of 20 μΙ_, samples can be spotted directly on the SRT fabricated swab and will be spotted on various substrates including but not limited to glass, plastic, metal, and leather followed by swab type wipe collection using the SRT swab device. Release of the captured human biological material can be triggered by the dissolution of SRT chemically by the use of chaotropic salt e.g 7.5 M G-HC1 and/or acidic pH that is compatible with DNA extraction. This will quantitatively release cells (with potential for >80% of bound material in less than 1 hr). Comparison with commercially available swabs (Cotton, Puritan Rayon and Foam, Copan 4N6 nylon, Fitzco polyester) can be performed to establish benchmarks.

[0038] It is expected that embodiments of the present disclosure will function to extract DNA and perform forensic type fragment amplification analysis. In this regard, DNA can be extracted initially using organic phenol, spin column and silica to demonstrate the efficient methods for SRT-based SCD. Quantification of DNA extracted can be performed by Quanitfiler (ABI/LifeTech/ThermoFisher). Further optimization can be performed using particular DNA extraction methods. We have demonstrated that SRT in powder form is insoluble in water and biological pH range and dissolves completely in the DNA extraction solutions of phenol and Guanidinium Hydrocholride (G-HC1).

[0039] Quantification of DNA can establish indirectly the release of biological material and thus of DNA from the SRT swab. Forensic type short tandem repeat DNA fragment analysis from extracted DNA can be performed using AmpliSTR amplification kit (ABI/LifeTech/ThermoFisher) to establish that the DNA extracted is amplifiable and compared to DNA extracted by the other commercially available swabs.

[0040] DNA quantification can be compared to direct DNA extraction of samples of

25K to 400K cells as 100% recovery and plotted with DNA recovery from various cell quantities. Similarly, relative fluorescence unit (RFU) and peak height ratio (PUR) can be calculated from STR amplifications, compared and data presented to establish reliable amplification and to exclude any inhibitory effect of the SRT swab SCD.

[0041] SRT foams: SRT proteins are rubbery solids above its glass transition temperature, and therefore, can be thermally shaped into any 3D geometry including thin films or foams. SRT foams are made by forming gas bubbles with the use of a blowing agent or using solution casting method with the use of salt. The physical properties of the foam is tunable due to molecular architecture of the structural SRT protein. For example, rigid foams can be manufactured by decreasing the molecular weight or increasing crystallinity percentage of the SRT proteins. Figure la shows an example of rigid foam that is highly cross-linked (40% crystallinity is characterized by X-Ray Diffraction in Figure lb) β-sheets with in the structure. Figure lc shows the porous structure of the protein foam. Data presented in Figure 1 were obtained using full length SRT from Loligo vulgaris.

[0042] SRT mechanical properties: SRT protein has excellent mechanical properties in both dry and wet conditions, exceeding most natural and synthetic polymers while having the unique capability of being thermally shaped into various functional forms (e.g., thin films to 3D printing). Both storage and loss modulus of SRT are constant as a function of frequency between 0.1 to 150 Hz. Stable frequency response is an important factor for mechanical performance of SCD devices in rotary devices during forensic sample

homogenization.

[0043] SCD's and SRT protein based SCD: Single nucleotide polymorphism (S P) studies are gaining prominence in clinical application of pharmacogenomics and especially related to drug interaction based on genotype. These require very little template DNA or RNA and thus amenable to quick swabbing technique for sample collection. The current disclosure includes DNA and/or RNA collection and extraction from minute quantities of biological samples and integration of a pestle type device for sample collection followed by processing in a 1.5 mL microcentrifuge tube for DNA or RNA extraction. These pestles (similar to swab type SCD's) are custom designed and fabricated in various configurations for efficient sample capture followed by concomitant nucleic acid extraction (see Figure 2). Various groove types were designed in the tip of the pestle to initially increase surface area of collection material and later for homogenization and extraction. These can be coated with a layer of fiber/foam sample collection material. Data in Figure 2 are obtained using an 18kDa SRT protein from Loligo vulgaris that has the sequence:

MAAKLITLLAVIALSNYAYALLPGLLGGYGYPAATTYRQTTHHGYGGLYGGLGYHY

PAATAVSHTTHHAPYGYGGLYGGLYGGLGYPAAASVSTVHHPVGYGGYGLGAYG

AYGLGYGLHYPAATAVSHTTHHAPYGYGGLYGGLYGGLGAVSTVSHGLGYGLHHP VGYAGYGLGAT AVSHTTHHAP YGGFGYGLY (SEQ ID NO : 1 ).

[0044] We tested SRT protein (in powder form) to ascertain the feasibility of sample collection matrix (Figure 2). Results demonstrate that SRT binds DNA (Figure 2a) under denaturing conditions and is triggered to release under non-denaturing low salt conditions, and the bound DNA is released with a predefined DNA elution trigger. Figure 2b

demonstrates that the SRT protein can be triggered to dissolve with quantitative release of all human cellular material that can be processed for DNA extraction using any of the common current procedures including organic (phenol) and G-HC1 as in used in spin column and silica beads (Figure 2c and 2d respectively). Only SRT completely dissolves in both G-HC1 and phenol, whereas all other commercially available swab were intact, including silk cocoon protein. Nylon slowly dissolves after 4 hrs became like a thick paste.

[0045] SRT protein fibers were obtained via electro-spinning (Figure 3a) and then spanned around a swab (Figure 3b) that could be made from wood, plastic or metal.

Alternatively, SRT is dip-coated on cotton swabs to create foam morphology (Figure 3c). This SCD devices can be used to capture blood samples (Figure 3d). Data in Figure 3 were obtained using an 22kDa SRT protein from Loligo vulgaris that has the sequence:

MVATVLVIMSMIAALSCQSEAALSVGTSVKTIRQSVHHGAVPAVGHTTVTHAVPHA YAYGGLPYGDAFGGLYGGLYGGLYGSPAATSVKTVSHGFHPTLPVGSTISHTTHGV HHPVTYGGLGLGGLGYGGLGYGGLGYGGLGYGGLGYGGLGYGGLGYGGLGAGGL YGLHYPGAVGLGYGLGGGYGGLYGLHLPAATSVSHTTHGVHHPALGLGLYGGAHL P AAS S VTHTTHGVAHPGLGLNYGVYGLH (SEQ ID NO:2).

[0046] While the disclosure has been described through specific embodiments, routine modifications will be apparent to those skilled in the art and such modifications are intended to be within the scope of the present invention.