Preservation of biological materials in non-aqueous fluid media
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Publication Number
US-9840731-B2
Publication Date
2017-12-12
Expiration Date
Abstract
The invention provides compositions and methods for preserving a biological material—such as a protein, a nucleic acid or a biological sample, or any combination thereof—in a substantially water-free, nonionic or ionic organic solvent. Improved preservation, including for example the stability and/or the solubility of the biological material in the substantially water-free fluid medium, is achieved with compositions comprising one or more substances (e.g., an antioxidant) described in the disclosure, and/or a metal salt. The biological material is soluble and stable, and retains its function and activity, when it is preserved in the substantially water-free fluid medium at ambient temperature or higher for extended periods of time. Therefore, the composition comprising the biological material does not need to be refrigerated or frozen during shipping or storage.
Core Innovation
The invention relates to a composition in a fluid state that is substantially free of water. The composition includes biological material comprising a polypeptide, a polynucleotide, a biological sample, or any combination thereof, configured to function as assay reagents, and the biological material retains its structural integrity, function, and/or retains its activity after storage.
The biological material is provided together with at least one solvent selected from an alcohol solvent and/or an ionic organic solvent, while maintaining substantially water-free conditions. The alcohol solvent comprises linear and branched C2-C6 acyclic alcohols having one or more hydroxyl groups and C3-C6 cyclic alcohols having one or more hydroxyl groups and three to six ring carbon atoms, optionally comprising one or more halide atoms.
For PCR and RT-PCR use, the composition includes reagents with a DNA polymerase and primer pairs for amplifying at least one nucleic acid locus, and optionally includes labeled forward and reverse primers. For RT-PCR use, the composition further includes a reverse transcriptase, reverse transcription primers, and primer pairs for amplifying complementary polydeoxyribonucleotide products, with optional dye labeling. For immunoassay use, the composition includes an antibody that is specific for a target antigen or analyte, with the antibody labeled with a dye or conjugated to a detection protein or enzyme, and optionally bound to a solid substrate.
Claims Coverage
The independent claim is clm-00001 and it defines a substantially water-free fluid composition with one or more assay reagent sets for PCR, RT-PCR, and/or immunoassays, together with a specified solvent system based on alcohol solvents or ionic organic solvents, while requiring retention of structural integrity, function, and/or activity after storage.
Substantially water-free fluid composition retaining biological material activity
A fluid composition comprising biological material and at least one solvent, wherein the composition is substantially free of water and the polypeptide, polynucleotide, or biological sample retains its structural integrity, function, and/or retains its activity after storage.
Alcohol solvent system for substantially water-free assay reagent compositions
The composition includes at least one alcohol solvent selected from linear and branched C2-C6 acyclic alcohols with one or more hydroxyl groups and C3-C6 cyclic alcohols with one or more hydroxyl groups and three to six ring carbon atoms, optionally comprising one or more halide atoms.
Ionic organic solvent system comprising organic salt and organic hydrogen bond donor
The composition includes at least one ionic organic solvent comprising an organic salt and an organic hydrogen bond donor.
PCR reagents with DNA polymerase and primer pairs in substantially water-free solvent
The composition includes one or more reagents for performing PCR comprising a DNA polymerase and at least one pair of a forward primer and a reverse primer for amplifying at least one nucleic acid locus, wherein the primer pair optionally is labeled with a dye.
RT-PCR reagents with reverse transcriptase, reverse transcription primers, and primer pairs
The composition includes one or more reagents for performing reverse transcription PCR (RT-PCR) comprising a reverse transcriptase, a DNA polymerase, at least one reverse transcription primer for reverse transcribing at least one polyribonucleotide to produce at least one polydeoxyribonucleotide complementary to the polyribonucleotide, and at least one pair of a forward primer and a reverse primer for amplifying the complementary polydeoxyribonucleotide, wherein the reverse transcription primer optionally is labeled with a dye and the forward and reverse primer pair optionally is labeled with a dye.
Immunoassay reagents with labeled or conjugated antibody and optional solid substrate
The composition includes one or more reagents for performing an immunoassay comprising an antibody specific for a target antigen or analyte, wherein the antibody is labeled with a dye or is conjugated to a detection protein or enzyme, and wherein the antibody optionally is bound to a solid substrate.
Overall, the claim coverage centers on a substantially water-free fluid composition that maintains biological material structural integrity, function, and/or activity after storage while incorporating assay reagent sets for PCR, RT-PCR, and/or immunoassays. The claim further specifies compatible solvent systems using either the recited alcohol solvents or ionic organic solvents comprising an organic salt and an organic hydrogen bond donor, with optional dye labeling and optional solid-substrate binding for immunoassays.
Stated Advantages
Retains structural integrity, function, and/or retains activity after storage.
Documented Applications
No documented applications found
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