Method and apparatus for in situ synthesis on a solid support

• US 6,589,726 B1
• Filed: 08/23/2000
• Issued: 07/08/2003
• Est. Priority Date: 09/04/1991
• Status: Expired due to Fees

• Current Assignee: Metrigenix Incorporated
• Original Assignee: Metrigenix Incorporated
• Inventors: Brennan, Thomas M., Butler, John H.
• Primary Examiner(s): Horlick, Kenneth R.
• Assistant Examiner(s): Spiegler, Alexander H.
• Application Number:
  US09/645,021
• Time in Patent Office: 1,049 Days
• Field of Search: 435/4, 435/6, 435/287.1, 435/287.8, 436/518, 436/524, 436/527, 427/2.11, 427/2.13
• US Class Current: 506/16
• 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 2219/00378 Piezoelectric or ink jet dispensers
• CPC Class Codes: B01J 2219/00432 Photolithographic masks
• CPC Class Codes: B01J 2219/00497 Features relating to the solid phase supports
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00529 DNA chips
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/00608 DNA chips
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00617 by chemical means
• CPC Class Codes: B01J 2219/00619 using hydrophilic or hydrophobic regions
• CPC Class Codes: B01J 2219/00621 by physical means, e.g. trenches, raised areas
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/0063 Other, e.g. van der Waals forces, hydrogen bonding
• CPC Class Codes: B01J 2219/00635 by reactive plasma treatment
• 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 2219/00675 : In-situ synthesis on the substrate
• CPC Class Codes: B01J 2219/00711 : Light-directed synthesis
• CPC Class Codes: B01J 2219/00722 : Nucleotides
• CPC Class Codes: B01J 2219/00725 : Peptides
• CPC Class Codes: B82Y 30/00 : Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C03C 17/30 : with silicon-containing compounds
• CPC Class Codes: C07B 2200/11 : Compounds covalently bound to a solid support
• CPC Class Codes: C07B 61/00 : Other general methods
• CPC Class Codes: C07H 21/00 : Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C07K 1/045 : using devices to improve synthesis, e.g. reactors, special vessels
• 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: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 40/10 : Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 50/14 : Solid phase synthesis, i.e. wherein one or more library building blocks are bound to a solid support during library creation; Particular methods of cleavage from the solid support
• CPC Class Codes: C40B 60/14 : for creating libraries

Abstract

The present invention relates to methods for fabricating solid supports. More specifically, the present invention features methods for fabricating solid supports for in situ synthesis and for carrying out large numbers of reactions. The present invention also features solid supports with in situ synthesized long polynucleotides.

Claims

1. A method for fabricating solid supports comprising the steps of:

(a) reacting a support surface with a first reagent to form a hydrophilic surface;

(b) coating said support surface with a photoresist substance;

(c) exposing selected regions of said support surface to light;

(d) developing said support surface to form a patterned exposed surface and photoresist coated surface;

(e) reacting the exposed surface with a second reagent to form hydrophobic sites; and

(f) removing the photoresist coat from said photoresist coated surface.

3. The method according to claim 1 or 2 wherein said support surface has about 1-10,000 hydrophilic sites per cm².

4. The method according to claim 1 or 2 wherein said support surface as about 10-500,000 hydrophilic sites.

5. The method according to claim 1 or 2 wherein the area of each hydrophilic site is about 0.1×

10<sup>−

5</sup> to 0.1 cm².

6. The method according to claim 1 or 2 wherein said support surface is a glass or silicon surface.

7. The method according to claim 1 wherein said first reagent is aminoalkylsilane or hydroxylalkylsilane.

9. The method according to claim 1, wherein said second reagent is fluoroalkylsiloxane or alkylsilane.

11. The method according to claim 1 or 2, wherein said hydrophilic sites are covalently or non-covalently attached to nucleic acids.

12. The method according to claim 1 or 2 wherein said first or second reagent is delivered using an ink-jet apparatus.

13. The method according to claim 1 or 2 wherein said first or second reagent is delivered using a piezoelectric pump.

2. A method for fabricating solid supports comprising the steps of:

(a) reacting a support surface with a first reagent to form a hydrophobic surface;

(b) coating said support surface with a photoresist substance;

(c) exposing selected regions of said support surface to light;

(d) developing said support surface to form a patterned exposed surface and photoresist coated surface;

(e) removing the photoresist coat from the photoresist coated surface; and

(f) reacting the exposed surface with a second reagent to form hydrophilic sites.

8. The method according to claim 2 wherein said first reagent is fluoroalkylsiloxane or alkylsilane.

10. The method according to claim 2, wherein said second reagent is aminoalkylsilane or hydroxylalkylsilane.