Surface with tethered polymeric species for binding biomolecules

• US 6,927,029 B2
• Filed: 12/03/2001
• Issued: 08/09/2005
• Est. Priority Date: 12/03/2001
• Status: Expired due to Fees

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Agilent Technologies, Inc.
• Inventors: Roitman, Daniel B., Lefkowitz, Steven M., Holcomb, Nelson R.
• Primary Examiner(s): FORMAN, BETTY J
• Application Number:
  US10/005,577
• Publication Number:
  US 20030104397A1
• Time in Patent Office: 1,345 Days
• Field of Search: 435/6, 435/7.1, 435/174, 435/283.1, 435/287.2, 435/287.9, 536/23.1, 536/24.3, 530/300, 530/350
• US Class Current: 435/6.11
• 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 33/545 Synthetic resin

Abstract

The invention provides a solid support for adsorbing a biomolecule. The support comprises a surface coating having a non-nucleotidic polymer tethered to a surface reactive site. The polymer comprises a backbone, terminus, and adsorbing moieties covalently attached to the backbone and capable of adsorbing a biomolecule that can assume a plurality of conformations. The polymer is generally tethered to the surface at its terminus and the backbone exhibits sufficient mobility and flexibility such that a biomolecule adsorbed by the adsorbing moieties can assume a desired conformation for hybridization. Also provided is a process for preparing a surface coating having a functionalized surface.

Claims

1. A process for preparing a solid support, comprising:

(a) providing a solid support comprising a surface coating having a surface reactive hydroxyl, carboxyl, amino or thiol group;

(b) contacting the surface reactive hydroxyl, carboxyl, amino or thiol group with a plurality of monomers;

(c) polymerizing said monomers to produce a solid support having a surface tethered polymer covalently linked to said surface coating, said surface tethered polymer having at least one adsorbing moiety for adsorbing a biomolecule; and

(d) linking a biomolecule to said polymer via said adsorbing moiety.

2. The process of claim 1, wherein a portion of said biomolecule is a linking moiety.

3. The process of claim 1, wherein said polymer is substantially linear.

4. The process of claim 1, wherein said adsorbing moiety is an amine group.

5. The process of claim 1, wherein said biomolecule comprises an oligonucleotide or polynucleotide.

6. The process of claim 1, further comprising polymerizing an additional non-nucleotidic polymer tethered to said surface coating, said non-nucleotidic polymer comprising additional adsorbing moieties for adsorbing additional biomolecules.

7. The method of claim 1, wherein at least one of said monomers reacts with said surface reactive hydroxyl, carboxyl, amino or thiol group to covalently bond said polymer to said surface coating.

8. A process for preparing a solid support capable of adsorbing a biomolecule, comprising:

(a) providing a solid support comprising a surface coating having a surface reactive hydroxyl, carboxyl, amino or thiol group;

(b) contacting the surface reactive hydroxyl, carboxyl, amino or thiol group with a plurality of monomers; and

(c) polymerizing said monomers to produce a solid support having a surface tethered polymer covalently linked to said surface coating, said surface tethered polymer having at least one adsorbing moiety for adsorbing a biomolecule,

wherein said polymer is a vinyl polymer.

9. The process of claim 8, wherein said vinyl polymer is a poly-(vinylamine).

10. A process for preparing a solid support containing a probe biomolecule capable of hybridization to a target species, comprising:

(a) providing a solid support comprising a surface coating having surface reactive hydroxyl, carboxyl, amino or thiol group,

(b) contacting the surface reactive hydroxyl, carboxyl, amino or thiol group with a plurality of monomers; and

(c) polymerizing said monomers to produce a solid support having a surface tethered polymer covalently linked to said surface coating, said surface tethered polymer having adsorbing sites for adsorbing biomolecules, wherein said surface tethered polymer is capable of assuming a plurality of conformations and exhibits sufficient mobility and flexibility such that the number of biomolecules adsorbed by the adsorbing moieties is maximized; and

(d) contacting the surface tethered polymer with the probe biomolecule.

11. The process of claim 10, wherein a portion of said biomolecule is a linking moiety.

12. The process of claim 11, wherein said vinyl polymer is a poly-(vinylamine).

13. The process of claim 10, wherein said polymer is substantially linear.

14. The process of claim 10, wherein said polymer is a vinyl polymer.

15. The process of claim 10, wherein said adsorbing moieties are amine groups.

16. The process of claim 10, wherein said biomolecule comprises an oligonucleotide or polynucleotide.

17. The process of claim 10, further comprising polymerizing a non-nucleotidic polymer tethered to said surface coating, said non-nucleotidic polymer comprising additional adsorbing moieties adapted to adsorb an additional biomolecule.

18. A process for preparing a solid support capable of adsorbing a biomolecule, comprising:

(a) providing a solid support comprising a surface coating having a surface reactive surface reactive hydroxyl, carboxyl, amino or thiol group;

(b) contacting the surface reactive hydroxyl, carboxyl, amino or thiol group with vinyl monomers; and

,

(c) polymerizing said monomers to produce a solid support having a surface tethered vinyl polymer covalently linked to said surface coating, said surface tethered vinyl polymer having at least one adsorbing moiety for adsorbing a biomolecule.

19. The process of claim 18, wherein said polymerization is done in the presence of cerium.

20. The process of claim 18, wherein said vinyl polymer is a poly-(vinylamine).