Derivatization of solid supports and methods for oligomer synthesis

• US 5,624,711 A
• Filed: 04/27/1995
• Issued: 04/29/1997
• Est. Priority Date: 04/27/1995
• Status: Expired due to Term

• Current Assignee: Affymetrix, Inc. (Thermo Fisher Scientific Incorporated)
• Original Assignee: Affymax Incorporated
• Inventors: Fujimoto, David, Sundberg, Steven A.
• Primary Examiner(s): Beck, Shrive
• Assistant Examiner(s): CAMERON, ERMA C
• Application Number:
  US08/431,196
• Time in Patent Office: 733 Days
• Field of Search: 436/518, 436/524-535, 530/333, 530/334, 530/335, 530/336, 530/337, 435/4, 435/7.1, 422/131, 422/134, 427/407.2, 427/387, 427/261, 427/287
• US Class Current: 506/32
• 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 2219/00364 Pipettes
• CPC Class Codes: B01J 2219/00378 Piezoelectric or ink jet dispensers
• CPC Class Codes: B01J 2219/0043 for direct application of reagents, e.g. through openings in a shutter
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/00504 Pins
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00529 DNA chips
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00592 Split-and-pool, mix-and-divide 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 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00641 the porous medium being continuous, e.g. porous oxide substrates
• 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: B05D 1/185 : applying monomolecular layers B05D1/204 takes precedence
• CPC Class Codes: B82Y 30/00 : Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: B82Y 40/00 : Manufacture or treatment of nanostructures
• CPC Class Codes: C03C 17/30 : with silicon-containing compounds
• CPC Class Codes: C03C 17/3405 : with at least two coatings of organic materials C03C17/36, C03C17/42 take precedence
• 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 60/14 : for creating libraries
• CPC Class Codes: C40B 80/00 : Linkers or spacers specially adapted for combinatorial chemistry or libraries, e.g. traceless linkers or safety-catch linkers
• CPC Class Codes: G01N 33/54353 : with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393

Abstract

Methods and derivatized supports which are useful in solid-phase synthesis of peptides, oligonucleotides or other small organic molecules as well as arrays of ligands. The methods provide means to control the functional site density on a solid support. Some of the derivatized supports are polymer-coated or glycan-coated. Other methods for regenerating the surface of a used ligand array are also provided.

Claims

1. A method for affixing functional sites to the surface of a solid substrate, comprising:

(a) contacting said solid substrate with a derivatizaton reagent, said reagent having a substrate attaching group and a reactive site, to covalently bind said substrate attaching group to said substrate and thereby form a derivatized substrate having reactive sites; and

(b) contacting said derivatized substrate with a mixture comprising linking molecules and diluent molecules, each of said linking molecules and diluent molecules having reactive groups capable of covalently binding to said reactive sites and said linking molecules alone further having a functional site for synthesis initiation, wherein said linking molecules and said diluent molecules are different and have different reactive properties and the ratio of said linking molecules to said diluent molecules in said mixture is selected to control the functional site density on said surface, to thereby bind said linking molecules and said diluent molecules to said substrate in accordance with said ratio.

2. A method in accordance with claim 1, wherein said substrate attaching group is a member selected from the group consisting of a trialkoxysilyl radical and a trichlorosilyl radical.

3. A method in accordance with claim 1, wherein said reactive site is a member selected from the group consisting of amino, hydroxyl, carboxylic acid, thiol, ester, amide, isocyanate and isothiocyanate.

4. A method in accordance with claim 1, wherein said derivatization reagent is an aminoalkyltrialkoxysilane.

5. A method in accordance with claim 1, wherein said functional site is a member selected from the group consisting of amino, hydroxyl, carboxylic acid, thiol, ester, amide, isocyanate and isothiocyanate.

6. A method in accordance with claim 1, wherein said diluent molecules are protected amino acids.

7. A method in accordance with claim 1, wherein said diluent molecules are members selected from the group consisting of protected glycine, protected serine and protected lysine, protected glutamic acid, protected aspartic acid, protected ornithine and protected phenylalanine.

8. A method in accordance with claim 1, wherein said ratio is from about 1:

2 to about 1;

200.