Reversible surface functionalization

• US 10,684,281 B2
• Filed: 05/03/2018
• Issued: 06/16/2020
• Est. Priority Date: 08/21/2014
• Status: Active Grant

• Current Assignee: Illumina Cambridge Limited (Illumina Incorporated)
• Original Assignee: Illumina Cambridge Limited (Illumina Incorporated)
• Inventors: von Hatten, Xavier, George, Wayne N., Richez, Alexandre, Dingwall, Anne-Cecile, Brown, Andrew A.
• Primary Examiner(s): Forman, Betty J
• Application Number:
  US15/970,702
• Publication Number:
  US 20180267034A1
• Time in Patent Office: 775 Days
• CPC Class Codes: C09D 183/04 Polysiloxanes
• CPC Class Codes: C12N 11/10 the carrier being a carbohydrate
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2563/131 the label being a member of a cognate binding pair, i.e. extends to antibodies, haptens, avidin
• CPC Class Codes: C12Q 2565/543 characterised by the use of two or more capture oligonucleotide primers in concert, e.g. bridge amplification
• CPC Class Codes: G01N 33/543 with an insoluble carrier for immobilising immunochemicals
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
• CPC Class Codes: G01N 33/551 the carrier being inorganic

Abstract

Some embodiments described herein relate to a substrate comprising a silane functionalized surface for reversibly immobilizing a biological molecule of interest, such as oligonucleotides, polynucleotides, or protein. Methods for immobilizing the biological molecule and the use in DNA sequencing and other diagnostic applications are also disclosed.

Claims

1. A substrate comprising a biological molecule immobilized to a surface of the substrate,

wherein the biological molecule is covalently bonded to a host molecule comprising a hydrophobic portion and a hydrophilic portion;

and wherein the host molecule is an optionally substituted cyclodextrin or a derivative thereof;

wherein the surface of the substrate comprises functionalized silane covalently attached thereto, and the functionalized silane comprises a guest moiety that can form a host-guest complex with the host molecule; and

wherein the biological molecule is immobilized through the host-guest complex between the functionalized silane and the host molecule.

2. The substrate of claim 1, wherein the guest moiety of the functionalized silane comprises a hydrophobic moiety.

3. The substrate of claim 2, wherein the host-guest interaction is a non-covalent interaction between the guest moiety of the functionalized silane and the host molecule.

4. The substrate of claim 3, wherein the host-guest non-covalent interaction is between the hydrophobic portion of the host molecule and the hydrophobic moiety of the functionalized silane.

5. The substrate of claim 2, wherein the hydrophobic moiety of the functionalized silane comprises aryl, heteroaryl, heterocyclyl, cycloalkyl, or cycloalkenyl, or optionally substituted variants thereof.

6. The substrate of claim 5, wherein the hydrophobic moiety comprise aryl.

7. The substrate of claim 5, wherein the hydrophobic moiety comprises cycloalkenyl.

8. The substrate of claim 7, wherein the cycloalkenyl is norbornene.

9. The substrate of claim 1, wherein the biological molecule comprises amino acid, peptide, nucleoside, nucleotide, oligonucleotide, polynucleotide, protein, sugar, or polysaccharide, or combinations thereof.

10. The substrate of claim 1, wherein the biological molecule is present on a cell, a virus or a particle, thereby immobilizing the cell, virus or particle through the host-guest interaction between the functionalized silane and the host molecule.

11. The substrate of claim 1, wherein the biological molecule is covalently bonded to the hydrophilic portion of the host molecule.

12. The substrate of claim 1, wherein the biological molecule comprises a oligonucleotide.

13. The substrate of claim 12, wherein the oligonucleotide is covalently bonded to the hydrophilic portion of the host molecule.

14. The substrate of claim 1, wherein the host molecule is β

-cyclodextrin.

15. The substrate of claim 1, wherein the surface of the substrate comprises glass.

16. The substrate of claim 1, wherein the guest moiety is substituted with one or more fluoro groups.

17. The substrate of claim 1, wherein the guest moiety of the functionalized silane is capable of forming covalent bonding with a functionalized polymer or hydrogel coating for surface functionalization.