Method of preparing a modified surface

• US 8,137,942 B2
• Filed: 10/31/2007
• Issued: 03/20/2012
• 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: Roitman, Daniel, Peluso, Paul, Foquet, Mathieu
• Primary Examiner(s): Naff, David M
• Application Number:
  US11/981,552
• Publication Number:
  US 20090061430A1
• Time in Patent Office: 1,602 Days
• Field of Search: None
• US Class Current: 435/176
• 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 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 reactive moiety, and a second surface modifying agent not coupled to the 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; and

coupling an enzyme to the reactive moiety on the surface.

2. The method of claim 1, wherein the reactive moiety comprises a specific binding group.

3. The method of claim 2 wherein the specific binding group is selected from avidin, streptavidin, biotin, binding peptides, antibodies, antigens, glutathione, and GST.

4. The method of claim 1 wherein the first and second surface modifying agents each comprise silanes.

5. The method of claim 1 wherein the first modifying agent is selected from epoxysilane, aminosilane, activated carboxylic acid silane, isocyanatosilane, aldehyde silane, mercaptosilane, vinyl silane, hydroxyterminated silane, and acrylate silane.

6. The method of claim 1 wherein the first and second surface modifying agents comprise PEG silanes.

7. The method of claim 6 wherein the first surface modifying agent comprises Silane-PEG-epoxide, and the second surface modifying agent comprises Silane-PEG-OH.

8. The method of claim 1 wherein the enzyme is coupled indirectly to the reactive moiety through an intermediate binding group.

9. The method of claim 8, wherein the enzyme comprises a nucleic acid polymerase.

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

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

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