Articles having localized molecules disposed thereon and methods of producing same

• US 10,655,172 B2
• Filed: 06/27/2014
• Issued: 05/19/2020
• 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:
  US14/318,324
• Publication Number:
  US 20140315756A1
• Time in Patent Office: 2,153 Days
• Field of Search: None
• 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 system comprising:

an array of nanoscale wells disposed through an opaque layer, each well having a bottom surface formed from a transparent layer underlying the opaque layer;

wherein a plurality of the wells in the array comprise an enzyme bound to the bottom surface and located in an observation region; and

wherein portions of the array other than the bottom surface of the wells are substantially free of the enzyme;

an illumination source that provides illumination to the observation regions through the transparent layer; and

an optical detector that receives electromagnetic radiation emitted through the transparent layer from the observation regions.

2. The system of claim 1, wherein the transparent layer comprises SiO₂.

3. The system of claim 1, wherein the opaque layer comprises a metal or metal oxide.

4. The system of claim 1, wherein the opaque layer comprises aluminum.

5. The system of claim 1, wherein the opaque layer is passivated with a compound comprising one or more phosphonic acid groups.

6. The system of claim 1, wherein the enzyme is a nucleic acid polymerase.

7. The system of claim 6, wherein the polymerase is part of a nucleic acid polymerase/template/primer complex.

8. The system of claim 7, wherein the wells comprise a solution comprising one or more nucleotides or nucleotide analogs.

9. The system of claim 1, wherein the plurality of the wells in the array comprise a single active enzyme molecule bound to the bottom surface.

10. The system of claim 1, wherein the enzyme is bound to the bottom surface through one or more silanes.

11. The system of claim 1, wherein the enzyme is bound to the bottom surface through a binding or coupling group applied to the bottom surface.

12. The system of claim 1, wherein the enzyme is bound to the bottom surface through biotin.

13. The system of claim 1, wherein the enzyme is bound to the bottom surface through a biotin-PEG-silane.