Arrays of protein-capture agents and methods of use thereof

• US 6,780,582 B1
• Filed: 05/17/2000
• Issued: 08/24/2004
• Est. Priority Date: 07/14/1998
• Status: Expired due to Fees

• Current Assignee: Zyomyx, Inc.
• Original Assignee: Zyomyx, Inc.
• Inventors: Wagner, Peter, Itin, Christian, Nock, Steffen, Ault-Riche, Dana
• Primary Examiner(s): Chin, Christopher L.
• Application Number:
  US09/574,692
• Time in Patent Office: 1,560 Days
• Field of Search: 435/6, 435/7.1, 435/287.1, 435/287.9, 435/288.3, 435/288.4, 435/287.2, 436/518, 436/524, 436/525, 436/527, 436/528, 436/532, 436/533, 436/535, 436/536, 422/57
• US Class Current: 506/32
• 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 2219/00619 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 2219/00635 by reactive plasma treatment
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00644 the porous medium being present in discrete locations, e.g. gel pads
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00702 Processes involving means for analysing and characterising the products
• 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: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C40B 40/10 Libraries containing peptides or polypeptides, or derivatives thereof
• 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
• CPC Class Codes: G01N 33/551 the carrier being inorganic

Abstract

Arrays of protein-capture agents useful for the simultaneous detection of a plurality of proteins which are the expression products, or fragments thereof, of a cell or population of cells in an organism are provided. A variety of antibody arrays, in particular, are described. Methods of both making and using the arrays of protein-capture agents are also disclosed. The invention arrays are particularly useful for various proteomics applications including assessing patterns of protein expression and modification in cells.

Claims

1. A biochip for displaying attached polynucleotides comprising:

(a) a substrate having an oxide or nitride surface; and

(b) an ordered hydrocarbon monolayer formed from first alkyl chains, said first alkyl chains having proximal and distal ends, said first alkyl chains interacting with each other to form said ordered hydrocarbon monolayer, said ordered hydrocarbon monolayer being attached to said surface through one or more silane residues on each of said first alkyl chains'"'"' proximal ends, said first alkyl chains'"'"' distal ends having a functional group capable of covalently attaching a polynucleotide to said first alkyl chain,

wherein the biochip further comprises one or more border regions separating two or more of said immobilization regions to form two or more spaced apart immobilization regions.

3. The biochip of claim 1 wherein said functional group is one of hydroxyl, carboxyl, amino, aldehyde, carbonyl, methyl, methylene, alkene, alkyne, carbonate, aryliodide, or vinyl groups.

4. The biochip of claim 1 wherein said functional group is a photoactivatable functional group.

5. The biochip of claim 1 wherein the first alkyl chains are the same length.

6. The biochip of claim 1 wherein the oxide or nitride surface is selected from the group consisting of silicon oxide titania, tantalum oxide, silicon nitride, indium tin oxide, magnesium oxide, alumina, quanz, glass, and silica.

7. The biochip of claim 1 wherein the first alkyl chains are from 8 to 22 carbons in length.

2. A biochip for displaying attached polynucleotide comprising:

(a) a substrate having an oxide or nitride surface; and

(b) an ordered hydrocarbon monolayer formed from first alkyl chains, said first alkyl chains having proximal and distal ends, said first alkyl chains interacting with each other to form said ordered hydrocarbon monolayer, said ordered hydrocarbon monolayer being attached to said surface through one or more silane residues an each of said first alkyl chains'"'"' proximal ends, said first alkyl chains'"'"' distal ends each having a functional group capable of covalently attaching a polynucleotide to said first alkyl chain,

wherein said ordered hydrocarbon monolayer is formed from a ratio of said first mentioned alkyl chains having a first mentioned functional group for attaching polynucleotides, and second alkyl chains, said second alkyl chains having proximal and distal ends, said alkyl chains interacting with each other to form said ordered hydrocarbon monolayer, said ordered hydrocarbon monolayer being attached to said surface through one or more second silane residues on each of said second alkyl chains'"'"' proximal ends, and

wherein said ordered hydrocarbon monolayer is made from two or more different kinds of molecules.