Reactive surfaces, substrates and methods of producing same

• US 20070077564A1
• Filed: 09/30/2005
• Published: 04/05/2007
• Est. Priority Date: 09/30/2005
• Status: Active Grant

• Current Assignee: Pacific Bioscience of California Incorporated (L'Oreal SA)
• Original Assignee: Pacific Bioscience of California Incorporated (L'Oreal SA)
• Inventors: Peluso, Paul, Roitman, Daniel, Foquet, Mathieu
• Granted Patent: US 7,763,423 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00628 Ionic
• CPC Class Codes: B01J 2219/0063 Other, e.g. van der Waals forces, hydrogen bonding
• 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: B01J 2219/00725 Peptides
• CPC Class Codes: G01N 33/543 with an insoluble carrier for immobilising immunochemicals
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• 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

Abstract

Reactive surfaces, substrates and methods of producing and using such substrates and surfaces are provided. The substrates and surfaces provide low density reactive groups preferably on an otherwise non-reactive surface for use in different applications including single molecule analyses.

Claims

1. A substrate comprising:

a surface comprising a first reactive moiety coupled thereto; and

wherein the reactive moiety is coupled to the surface at a density of between about 1 reactive moiety per 50,000 nm² and 1 reactive moiety per 100 nm².

2. The substrate of claim 1, wherein the reactive moiety is coupled to the surface at a density of between about 1 reactive moiety per 8000 nm² and 1 reactive moiety per 300 nm²

3. The substrate of claim 1, wherein the reactive moiety is coupled to the surface at a density of between about 1 reactive moiety per 8000 nm² and 1 reactive moiety per 1000 nm²

4. The substrate of claim 1, wherein a remainder of the surface other than the reactive moieties is a substantially non-reactive surface.

5. The substrate of claim 1, wherein the reactive moiety is coupled to the surface through a linker group.

6. The substrate of claim 5, wherein the linker group comprises a polymer.

7. The substrate of claim 5, wherein the linker group comprises polyethylene glycol.

8. The substrate of claim 4, wherein the reactive moiety is coupled to the surface through a linker group, and the substantially non-reactive surface comprises the linker group without the reactive moiety.

9. The substrate of claim 1, wherein the first reactive moiety comprises a binding moiety.

10. The substrate of claim 9, wherein the binding moiety comprises a nonspecific binding moiety.

11. The substrate of claim 9, wherein the binding moiety comprises a specific binding moiety.

12. The substrate of claim 11, wherein the binding moiety comprises one member of a specific binding pair.

13. The substrate of claim 9, wherein the binding moiety is selected from the group of consisting of an antigen, an antibody, an binding fragment of an antibody, a polynucleotide, a binding peptide, biotin, avidin and streptavidin.

14. The substrate of claim 1, wherein the first reactive moiety comprises a catalytic moiety.

15. The substrate of claim 14, wherein the catalytic moiety comprises a catalytic metal.

16. The substrate of claim 14, wherein the catalytic moiety comprises an enzyme.

17. The substrate of claim 16, wherein the enzyme is selected from a nucleic acid polymerase, a ligase, a nuclease, a protease, a kinase and a phosphatase.

18. The substrate of claim 16, wherein the enzyme comprises a DNA polymerase.

19. The substrate of claim 1, wherein the surface comprises an observation area.

20. The substrate of claim 1, wherein the surface comprises an observation surface of an optical confinement.

21. The substrate of claim 19, wherein the observation area comprises the observation surface of a zero mode waveguide.

22. The substrate of claim 1, wherein the surface of the substrate comprises silica.

23. The substrate of claim 1, wherein the surface of the substrate is selected from glass, quartz, fused silica, and silicon.

24. A device, comprising:

a substrate having at least a first observation area provided therein, the observation area having an area of from about 100 nm² to 50000 nm²; and

from 1 to 3 reactive moieties coupled to the surface within the observation area.

25. The device of claim 24, wherein the observation area is bounded by an optically opaque cladding.

26. The device of claim 24, wherein the reactive moieties comprise an enzyme.

27. The device of claim 26, wherein the enzyme comprises a DNA polymerase.

28. A method of preparing a modified surface, comprising:

providing a substrate having a first reactive surface;

providing a mixture of first and second surface modifying agents, wherein the first and second surface modifying agents are each capable of coupling to the reactive surface, and are present in the mixture at a first ratio selected so that the first and second surface modifying agents couple to the reactive surface at a second ratio;

contacting the reactive surface with the mixture to produce the modified surface having first and second modifying agents coupled thereto at the second ratio.

29. A method of preparing a modified surface, comprising:

providing a surface to be modified;

contacting the surface to be modified with a surface modifying composition, wherein the surface modifying composition comprises;

a first surface modifying agent coupled to a desired reactive moiety, and a second surface modifying agent not coupled to the desired reactive moiety; and

the first surface modifying agent and second surface modifying agent are present in the surface modifying composition at a ratio that produces the modified surface, wherein the reactive moieties are present on the modified surface at a density of between about 1 reactive moiety per 50,000 nm² and 1 reactive moiety per 100 nm²

30. An apparatus, comprising a surface having a reactive moiety coupled thereto, wherein the reactive moiety is coupled to the surface at a density of less than 1 reactive moiety per 100 nm².

31. The apparatus of claim 30, wherein the reactive moiety is coupled to the surface at a density of less than 1 reactive moiety per 1000 nm²

32. The apparatus of claim 30, wherein the reactive moiety is coupled to the surface at a density of less than 1 reactive moiety per 10000 nm²

33. The apparatus of claim 30, wherein the reactive moiety comprises a molecular binding group.

34. The apparatus of claim 33, wherein the molecular binding group comprises a specific molecular binding group.

35. The apparatus of claim 34, wherein the specific molecular binding group is selected from an antibody, an antigen, avidin, streptavidin, biotin, a polynucleotide, a polynucleotide analog, a binding peptide.

36. A method of configuring a surface to provide a desired density of reactive moieties thereon, comprising treating the surface with a composition that substantially uniformly couples to the surface, the composition comprising a first component that does not contain the reactive moiety and a second component that comprises the reactive moiety, the second component being present in the composition at a concentration relative to the first component so as to provide the reactive moieties on the surface at the desired density.