Conjugation of carboxyl functional hydrophilic beads

Patent metadata

Field	Value
Publication number	US-10144968-B2
Application number	US-201615200588-A
Country	US
Kind code	B2
Filing date	Jul 1, 2016
Priority date	Jul 2, 2015
Publication date	Dec 4, 2018
Grant date	Dec 4, 2018

Abstract

A polymer substrate, such as a polymer coating or a polymer hydrogel network, includes carboxyl moieties that can be used as conjugation sites to which receptor or analyte molecules can be attached. In an example, the polymer substrate includes a polyacrylamide polymer network having alkanoic acid moieties or derivatives thereof, which can react with carboxyl activating compounds to provide an activated alkanoate moieties on the polyacrylamide network. Amine-terminated nucleic acids can react with the activated alkanoate moieties to capture the nucleic acid to the polymer network through an alkylamide moiety.

First claim

What is claimed is:

1. A method of preparing a polymeric substrate, the method comprising:
in a non-aqueous solvent, contacting the polymer substrate with a carboxyl activating compound, the polymer substrate including an alkanoic acid moiety or ester derivative thereof, the carboxyl activating compound and the alkanoic acid moiety reacting to form an activated alkanoate moiety; and
contacting a liphophilic amine-terminated oligonucleotide and the polymer substrate in the non-aqueous solvent, an amine termination moiety of the lipophilic amine-terminated oligonucleotide forming an alkanamide moiety in place of the activated alkanoate moiety.

2. The method of claim 1 , wherein the polymer substrate includes polyacrylamide with the alkanoic acid moiety.

3. The method of claim 2 , wherein the polyacrylamide is a crosslinked polyacrylamide.

4. The method of claim 2 , wherein the polyacrylamide further includes an alkanol moiety.

5. The method of claim 1 , wherein the alkanoic acid moiety has 2 to 8 carbons.

6. The method of claim 5 , wherein the alkanoic acid moiety has 3 to 6 carbons.

7. The method of claim 1 , wherein the alkanoic acid moiety is appended to an acrylamide functionality of the polyacrylamide by a polar moiety.

8. The method of claim 7 , wherein the polar moiety is a polyether structure.

9. The method of claim 8 , wherein the polyether structure includes 1 to 10 ether units.

10. The method of claim 9 , wherein the ether units are derived from ethylene oxide or propylene oxide units.

11. The method of claim 1 , wherein the polymer substrate is a polymer coating.

12. The method of claim 1 , wherein the polymer substrate is a polymer hydrogel network.

13. The method of claim 1 , wherein the carboxylate activating compound is a succinimidyl compound.

14. The method of claim 13 , wherein the succinimidyl compound includes a succinimidyl uronium compound, a succinimidyl phosphonium compound, or a combination thereof.

15. The method of claim 14 , wherein the succinimidyl compound is a succinimidyl uronium compound.

16. The method of claim 15 , wherein the succinimidyl uronium compound is an O-type uronium compound.

17. The method of claim 1 , further comprising contacting a template nucleic acid with the oligonucleotide, at least a portion of the template nucleic acid complementary to the oligonucleotide.

18. The method of claim 17 , further comprising extending the oligonucleotide complementary to the template nucleic acid.

19. The method of claim 1 , wherein the amine-terminated oligonucleotide includes the amine termination on a 5 ′ end.

20. The method of claim 1 , wherein the amine termination of the amine-terminated oligonucleotide includes an alkylamine termination moiety.

Assignees

• Life Technologies Corp
• Life Tech As

Inventors

• Fonnum Geir
• Menchen Steven M
• Lui Alfred
• Xiang Guobing
• Larsen Synne

Classifications

• C08J7/14

  with acids, their salts or anhydrides · CPC title

• C12Q1/6869 Primary

  Methods for sequencing · CPC title

• C12Q1/6853

  using modified primers or templates · CPC title

• C12Q1/6876 Primary

  Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes · CPC title

• C12Q1/6874 Primary

  involving nucleic acid arrays, e.g. sequencing by hybridisation · CPC title