Articles having localized molecules disposed thereon and methods of producing same

• US 8,772,202 B2
• Filed: 06/04/2012
• Issued: 07/08/2014
• Est. Priority Date: 03/30/2006
• Status: Active Grant

• Current Assignee: Pacific Bioscience of California Incorporated (L'Oreal SA)
• Original Assignee: Pacific Bioscience of California Incorporated (L'Oreal SA)
• Inventors: Rank, David R., Wegener, Jeffrey, Korlach, Jonas, Roitman, Daniel, Xu, Yue, Lyle, John, Turner, Stephen, Peluso, Paul, Otto, Geoff, Cicero, Ronald L.
• Primary Examiner(s): Gross, Christopher M
• Application Number:
  US13/487,978
• Publication Number:
  US 20120302459A1
• Time in Patent Office: 764 Days
• Field of Search: None
• US Class Current: 506/43
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00427 using masks
• CPC Class Codes: B01J 2219/0045 using optical fibres
• 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 2219/00617 by chemical means
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00635 by reactive plasma treatment
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00639 the compounds being trapped in or bound to a porous medium
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00711 Light-directed synthesis
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B82Y 20/00 Nanooptics, e.g. quantum optics or photonic crystals
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C12Q 1/6874 involving nucleic acid arrays, e.g. sequencing by hybridisation
• CPC Class Codes: C12Q 2527/127 the enzyme inhibitor or activator used
• CPC Class Codes: C12Q 2565/629 being a microfluidic device
• CPC Class Codes: Y10T 29/49002 : Electrical device making

Abstract

Methods of producing substrates having selected active chemical regions by employing elements of the substrates in assisting the localization of active chemical groups in desired regions of the substrate. The methods may include optical, chemical and/or mechanical processes for the deposition, removal, activation and/or deactivation of chemical groups in selected regions of the substrate to provide selective active regions of the substrate.

Claims

1. A method of preparing a substrate for selective immobilization of an enzyme thereon, comprising:

providing a substrate having a first surface component and a second surface component where the first and second surface components have different surface characteristics, wherein the substrate comprises a layer of the second surface component on a layer of the first surface component, and wherein the substrate comprises a zero mode waveguide disposed through the second surface component layer to the first surface component layer and exposing a portion of the first surface component layer, the portion of the first surface component layer thereby exposed forming a bottom reading surface of the zero mode waveguide; and

contacting the substrate with a composition that selectively associates with the second surface component, which composition comprises a compound comprising one or more phosphonic acid groups, thereby passivating the second surface component and preparing the substrate for selective immobilization of the enzyme atop the bottom reading surface in the zero mode waveguide.

2. The method of claim 1, wherein the first surface component comprises SiO₂ and the second surface component comprises a metal or metal oxide.

3. The method of claim 1, wherein the differing surface characteristics comprise surface charge.

4. The method of claim 3, wherein the first surface component has a negative surface charge and the second surface component has a positive surface charge.

5. The method of claim 1, comprising contacting the substrate with a silane that associates with the first surface component.

6. The method of claim 5, wherein the substrate is contacted with the composition that selectively associates with the second surface component and then contacted with the silane.

7. The method of claim 1, comprising selectively coupling the enzyme atop the bottom reading surface in the zero mode waveguide.

8. The method of claim 6, wherein selectively coupling the enzyme atop the bottom reading surface comprises contacting the substrate with poly(L-lysine)-poly(ethylene glycol) or poly(L-lysine)-poly(ethylene glycol)-biotin that associates with the first surface component and coupling the enzyme to the poly(L-lysine)-poly(ethylene glycol) or poly(L-lysine)-poly(ethylene glycol)-biotin.

9. The method of claim 1, wherein the composition that selectively associates with the second surface component is polyvinylphosphonic acid.

10. The method of claim 1, wherein the composition that selectively associates with the second surface component is selected from the group consisting of:

11. The method of claim 1, wherein the composition that selectively associates with the second surface component is selected from the group consisting of:

2-carboxyethyl phosphonic acid;

amino tri(methylene phosphonic acid);

1-hydroxyethylidene-1,1,-diphosphonic acid;

hexamethylenediaminetetra(methylenephosphonic acid);

diethylenetriamine penta(methylene phosphonic acid);

ethylenediamine tetra(methylene phosphonic acid);

bis(hexamethylene triamine penta(methylenephosphonic acid));

2-phosphonobutane-1,2,4-tricarboxylic acid; and

monoethanloamine diphosphonate.

12. The method of claim 1, wherein the composition that selectively associates with the second surface component comprises an alkyl phosphonate.

13. The method of claim 1, wherein the composition that selectively associates with the second surface component is selected from the group consisting of:

octyl phosphonic acid, decyl phosphonic acid, dodecyl phosphonic acid, hexadecyl phosphonic acid, octadecyl phosphonic acid, docosyl phosphonic acid, hydroxy-dodecyl phosphonic acid, hydroxy-undecenyl-phosphonic acid, and decanediylbis(phosphonic acid).

14. The method of claim 1, wherein the composition that selectively associates with the second surface component comprises a second coupling group.

15. The method of claim 7, wherein the enzyme comprises a polymerase.

16. A method of analyzing an enzyme that is selectively immobilized on a substrate, comprising:

a) selectively immobilizing the enzyme on the substrate by;

providing a substrate having a first surface component and a second surface component where the first and second surface components have different surface characteristics, wherein the substrate comprises a layer of the second surface component on a layer of the first surface component, and wherein the substrate comprises a zero mode waveguide disposed through the second surface component layer to the first surface component layer,

contacting the substrate with a composition that selectively associates with the second surface component, which composition comprises a compound comprising one or more phosphonic acid groups, and

selectively coupling the enzyme to the first surface component in the zero mode waveguide; and

b) after selectively immobilizing the enzyme on the substrate, providing illumination to and receiving electromagnetic radiation from an observation volume of the zero mode waveguide through the first surface component layer.

17. The method of claim 16, wherein the first surface component comprises SiO₂ and the second surface component comprises a metal or metal oxide.

18. The method of claim 16, wherein selectively coupling the enzyme to the first surface component comprises contacting the substrate with a silane that associates with the first surface component and coupling the enzyme to the silane.

19. The method of claim 18, wherein the silane comprises a first coupling group, and wherein selectively coupling the enzyme to the first surface component comprises coupling the enzyme to the first coupling group.

20. The method of claim 18, wherein the substrate is contacted with the composition that selectively associates with the second surface component and then contacted with the silane.

21. The method of claim 16, wherein the composition that selectively associates with the second surface component is polyvinylphosphonic acid.

22. The method of claim 16, wherein the enzyme comprises a polymerase.