Reactive surfaces, substrates and methods of producing and using same

• US 20090061430A1
• Filed: 10/31/2007
• Published: 03/05/2009
• 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 8,137,942 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

1-27. -27. (canceled)

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.

37. The method of claim 28, wherein the first surface modifying agent comprises a desired reactive moiety coupled thereto, such that the first surface modifying agent coupled to the first surface provides the desired reactive moiety upon the modified surface.

38. The method of claim 28, further comprising contacting the modified surface with a molecule of interest capable of coupling to only one of the first and second surface modifying agents.

39. The method of claim 38, wherein the molecule of interest is selected from a protein, a nucleic acid, and a specific binding group.

40. The method of claim 38, wherein the molecule of interest comprises a specific binding group selected from avidin, streptavidin, neutravidin and biotin.

41. The method of claim 28, wherein the first and second surface modifying agents each comprise silanes.

42. The method of claim 28, wherein the first and second surface modifying agents are independently selected from epoxysilane, aminosilane, activated carboxylic acid silane, isocyanatosilane, aldehyde silane;

mercaptosilane, vinyl silane, hydroxyterminated silane, and acrylate silane.

43. The method of claim 41, wherein the first and second surface modifying agents comprise PEG silanes.

44. The method of claim 43, wherein the first surface modifying agent comprises Silane-PEG-epoxide, and the second surface modifying agent comprises Silane-PEG-OH.

45. The method of claim 28, further comprising coupling an active enzyme to only one of the first and second surface modifying agents.

46. The method of claim 45, wherein the enzyme is coupled indirectly to the first or second surface modifying agent through an intermediate binding group.

47. The method of claim 46, wherein the enzyme comprises a nucleic acid polymerase.

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.

48. The method of claim 29, wherein the reactive moieties are present on the modified surface at a density of less than 1 per 1000 nm².

49. The method of claim 29, wherein the reactive moieties are present on the modified surface at a density of less than 1 per 10,000 nm².

50. The method of claim 29, wherein the reactive moieties are present on the modified surface at a density between about 1/2500 nm² and about 1/300 nm².

51. The method of claim 29, further comprising coupling a catalytic moiety to the reactive moiety on the modified surface.

52. The method of claim 51, wherein the catalytic moiety comprises a nucleic acid polymerase enzyme.

30-36. -36. (canceled)