Modified surfaces

• US 10,385,335 B2
• Filed: 12/05/2014
• Issued: 08/20/2019
• Est. Priority Date: 12/05/2013
• Status: Active Grant

• Current Assignee: Centrillion Technology Holdings Corporation (Centrillion Technology, Inc.)
• Original Assignee: Centrillion Technology Holdings Corporation (Centrillion Technology, Inc.)
• Inventors: McGall, Glenn
• Primary Examiner(s): Priest, Aaron A
• Application Number:
  US15/101,168
• Publication Number:
  US 20160298110A1
• Time in Patent Office: 1,719 Days
• Field of Search: None
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: C08F 220/56 Acrylamide; Methacrylamide
• CPC Class Codes: C08F 220/58 containing oxygen in addition to the carbonamido oxygen , e.g. N-methylolacrylamide, N-(meth)acryloylmorpholine
• CPC Class Codes: C09D 4/00 Coating compositions, e.g. paints, varnishes or lacquers, based on organic non-macromolecular compounds having at least one polymerisable carbon-to-carbon unsaturated bond ; Coating compositions, based on monomers of macromolecular compounds of groups C09D183/00 - C09D183/16
• CPC Class Codes: C12N 15/1093 General methods of preparing gene libraries, not provided for in other subgroups
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 1/6874 involving nucleic acid arrays, e.g. sequencing by hybridisation
• CPC Class Codes: C12Q 2523/101 Crosslinking agents, e.g. psoralen
• CPC Class Codes: C12Q 2527/153 Viscosity
• CPC Class Codes: C12Q 2531/113 PCR
• CPC Class Codes: C40B 50/18 using a particular method of attachment to the solid support

Abstract

Provided herein are methods and compositions for coating surfaces with polymers. The methods and compositions are suited for conducting biological reactions.

Claims

1. A method for performing an enzymatic reaction, comprising:

(a) providing a substrate comprising surface-bonded initiator species comprising a silane;

(b) conducting surface initiated polymerization of a mixture of (i) first monomers comprising acrylamide and/or ethoxylated acrylamide, each of which has no biomolecule attached to, and (ii) second monomers comprising acrydite, each of which has a biomolecule attached to, wherein said surface initiated polymerization starting from said initiator species, and coupling a plurality of said biomolecules to a polymer brush coating during said surface initiated polymerization, thereby producing said polymer brush coating with said plurality of said biomolecules coupled to said polymer brush coating; and

(c) performing one or more enzymatic reactions with said biomolecules on said substrate;

wherein the biomolecules are selected from the group consisting of;

oligonucleotides and polynucleotides, wherein the one enzymatic reaction is selected from the group consisting of;

polymerase chain reaction, sequencing reaction, sequencing by synthesis reaction, ligation reaction, extension reaction, and transcription reaction.

2. The method of claim 1, further comprising applying heat to said substrate.

3. The method of claim 1, wherein said polymer brush coating coupled with said plurality of biomolecules exhibits robustness wherein at least 90% of said biomolecules are retained on said surface after 40 cycles of sequencing by synthesis reactions.

4. The method of claim 1, wherein the substrate comprises at least 1,000,000 different types of biomolecules, and wherein each biomolecule is an oligonucleotide.

5. The method of claim 4, wherein said enzymatic reaction is an extension reaction.

9. The method of claim 1, wherein said polymer brush coating coupled with said plurality of biomolecules exhibits robustness in said polymerase chain reactions and wherein said polymerase chain reactions comprise the following reaction conditions:

(a) a denaturation step at a temperature of at least 85°

C. for at least 15 seconds;

(b) an annealing step at a temperature of at least 50°

C. for at least 15 seconds; and

(c) an extension step at a temperature of at least 70°

C. for at least 30 seconds.

10. The method of claim 1, wherein said first monomers comprise acrylamide.

11. The method of claim 10, wherein said first monomers further comprise N-(2-hydroxyethyl)acrylamide.

12. The method of claim 1, wherein said surface-bonded initiator species comprises 2-bromo-2-methyl-N,N-bis-(3-trimethoxysilanylpropyl)propionamide.

13. The method of claim 1, wherein said surface initiated polymerization comprises atom-transfer radical polymerization (ATRP).

14. The method of claim 1, wherein said second monomers are 5′

acrydite modified oligonucleotides or polynucleotides.

15. The method of claim 1, wherein said first monomers comprise ethoxylated acrylamide.

17. The method of claim 1, wherein robustness of said polymer brush coating coupled with said plurality of biomolecules is exhibited in said polymerase chain reaction:

at least 90% of said biomolecules are retained on said surface after 25 cycles of said polymerase chain reactions.

6. A method for making a modified surface, comprising:

(a) providing a surface;

(b) covalently bonding initiator species to said surface, wherein said initiator species comprises a silane; and

(c) conducting surface initiated polymerization of a mixture of (i) first monomers comprising acrylamide and/or ethoxylated acrylamide, each of which has no biomolecule attached to, and (ii) second monomers comprising acrydite, each of which has a biomolecule attached, wherein said surface initiated polymerization starting polymer from said initiator species, thereby producing a polymer brush coating comprising a plurality of polymer chains, each of which has said biomolecule attached.

7. The method of claim 6, wherein said initiator species comprises the molecule shown below:

8. The method of claim 6, wherein said second monomers are 5′

acrydite modified oligonucleotides.

16. The method of claim 6, wherein said first monomers comprise ethoxylated acrylamide.