Substrates and associated methods for elution of nucleic acids

Patent metadata

Field	Value
Publication number	US-9534214-B2
Application number	US-201314068633-A
Country	US
Kind code	B2
Filing date	Oct 31, 2013
Priority date	Oct 31, 2013
Publication date	Jan 3, 2017
Grant date	Jan 3, 2017

Abstract

A solid substrate for biological sample storage under dry-state and elution of biomolecules is provided. The dry, solid substrate is coated with saccharides, such as monosaccharides, oligosaccharides, polysaccharides or combinations thereof, and the substrate is comprised of one or more protein denaturing agents impregnated therein under a substantially dry state. A method for elution of biomolecules from biological samples is also provided. The compositions disclosed herein provide for enhanced elution and recovery of biomolecules, such as nucleic acids, from the sample. The sample is disposed on a substrate, dried to a substantially dry state; eluted from the biological sample dried on the substrate by rehydrating the substrate in an elution buffer.

First claim

The invention claimed is:

1. A saccharide coated substrate for eluting biomolecules from a biological sample, consisting essentially of:
a cellulose membrane;
a coating of saccharides disposed on the cellulose membrane, wherein saccharides in the coating of saccharides are selected from the group consisting of melezitose, raffinose, and a combination thereof; and
one or more protein denaturing agents impregnated in the cellulose membrane having a water content of less than 2%,
wherein the saccharide coated substrate is a non-water dissolvable material.

2. The saccharide coated substrate of claim 1 , wherein the cellulose membrane is a porous cellulose membrane.

3. The saccharide coated substrate of claim 1 , wherein the protein denaturing agents comprise salts, detergents, chaotropes, or combinations thereof.

4. The saccharide coated substrate of claim 3 , wherein the protein denaturing agents are selected from thiocyanate salts.

5. The saccharide coated substrate of claim 4 , wherein the protein denaturing agent is guanidinium thiocyanate.

6. The saccharide coated substrate of claim 3 , wherein the protein denaturing agents are selected from the group consisting of guanidinium hydrochloride, arginine, sodium dodecyl sulfate (SDS), urea, and combinations thereof.

7. The saccharide coated substrate of claim 1 , further comprising a reducing agent, a buffer, an anti-oxidant, a chelating agent, or combinations thereof.

8. The saccharide coated substrate of claim 7 , wherein the reducing agent is selected from the group consisting of dithiothreitol (DTT), 2-mercaptoethanol (2-ME), tris(2-carboxyethyl)phosphine (TCEP), and combinations thereof.

9. The saccharide coated substrate of claim 7 , wherein the buffer is selected from the group consisting of 2-amino-2-hydroxymethyl-propane-1,3-diol (Tris), 2-(N-morpholino) ethanesulfonic acid (MES), 3-(N-morpholino) propanesulfonic acid (MOPS), citrate buffers, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), phosphate buffers, and combinations thereof.

10. The saccharide coated substrate of claim 7 , wherein the anti-oxidant is selected from the group consisting of hydroquinone monomethyl ether (MEHQ), hydroquinone (HQ), toluhydroquinone (THQ), ascorbic acid, uric acid, and combinations thereof.

11. The saccharide coated substrate of claim 7 , wherein the chelating agent is selected from the group consisting of ethylenediaminetetraacetic acid (EDTA), citric acid, ethylene glycol tetraacetic acid (EGTA), and combinations thereof.

12. A method for eluting a protein, peptide or a nucleic acid from a biological sample disposed on a saccharide-coated solid substrate, consisting essentially of:
A cellulose membrane,
A coating of saccharides disposed on the cellulose membrane, wherein saccharides in the coating of saccharides are selected from the group consisting of melezitose, raffinose and combination thereof; and
one or more protein denaturing agents impregnated in the cellulose membrane, drying the biological sample to a water content of less than 2%,
contacting the biological sample to the substrate,
eluting the protein, peptide or nucleic acid from the biological sample dried on the substrate by rehydrating the substrate in an elution buffer.

13. The method of claim 12 , wherein the substrate further comprises a reducing agent, a buffer, an anti-oxidant, a chelating agent or combinations thereof impregnated therein.

14. The method of claim 12 wherein the biomolecule is DNA and the protein denaturing agent is one or more thiocyanate salts.

Assignees

• Gen Electric

Inventors

• Li Bing
• Grossmann Gregory Andrew
• Kvam Erik Leeming
• Bales Brian Christopher
• Davis Jason Louis

Classifications

• B01J20/22

  comprising organic material · CPC title

• B01J20/261

  obtained by reactions only involving carbon to carbon unsaturated bonds (macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds per se C08F) · CPC title

• B01J20/3251

  comprising at least two different types of heteroatoms selected from nitrogen, oxygen or sulphur · CPC title

• B01J20/3212

  consisting of a polymer obtained by reactions otherwise than involving only carbon to carbon unsaturated bonds · CPC title

• B01J20/3248

  the functional group or the linking, spacer or anchoring group as a whole comprising at least one type of heteroatom selected from a nitrogen, oxygen or sulfur, these atoms not being part of the carrier as such · CPC title