Protein arrays for high-throughput screening

• US 6,406,921 B1
• Filed: 07/14/1998
• Issued: 06/18/2002
• Est. Priority Date: 07/14/1998
• Status: Expired due to Term

• 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:
  US09/115,455
• Time in Patent Office: 1,435 Days
• Field of Search: 435/287.9, 435/286.1, 435/287.1, 435/288.3, 435/288.4, 435/7.1, 427/261, 427/287, 427/387, 427/407.2, 427/134, 427/518, 436/518, 436/524, 436/525, 436/527, 436/528, 436/532, 436/533, 436/535, 436/536
• US Class Current: 436/518
• 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 2219/0063 Other, e.g. van der Waals forces, hydrogen bonding
• CPC Class Codes: B01J 2219/00635 by reactive plasma treatment
• CPC Class Codes: B01J 2219/00637 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 2219/00702 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: C07K 2319/20 containing a tag with affinity for a non-protein ligand
• 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
• CPC Class Codes: G01N 33/6845 : Methods of identifying protein-protein interactions in protein mixtures

Abstract

Protein arrays and protein-coated substrates for the parallel, in vitro screening of biomolecular activity are provided. Methods of using the protein-coated substrates and protein arrays are also disclosed. A plurality of different members of a single protein family may be immobilized on the protein-coated substrate or array. The protein-coated substrates and protein arrays are particularly useful in high-throughput drug screening and clinical diagnostics.

Claims

1. An array device comprising:

a substrate defining a surface;

an array of spaced-apart protein immobilization regions over said surface, said protein immobilization regions being effective to immobilize one or more selected proteins, said immobilization regions being resistant non-specific protein binding after immobilization of said one or more selected proteins,

one or more hydrophilic border regions surrounding each protein immobilization region and separating such protein immobilization regions from one another, said border regions each comprising (i) an ordered hydrophobic monolayer formed of alkyl chains having proximal ends which are chemisorbed or physisorbed to said surface within said protein immobilization regions, and opposite hydrophobic distal ends, (ii) a hydrophilic monolayer attached to said hydrophobic monolayer, said hydrophilic monolayer comprising hydrophilic chains, each hydrophilic chain having a proximal end by which said hydrophilic chain is linked to an alkyl chain distal end, and an opposite hydrophilic distal end, together said hydrophobic monolayer and hydrophilic monolayers forming said hydrophilic border regions which are effective to resist non-specific protein binding,

wherein application of one or more of said selected proteins to selected protein immobilization regions is effective to form an array of protein regions having one or more selected proteins displayed within said protein immobilization regions such that each protein immobilization region is separated from other protein immobilization regions by hydrophilic border regions resistant to non-specific protein binding to form a protein array resistant to non-specific binding of proteins.

2. The device of claim 1, wherein said hydrophobic polymer chains are hydrocarbon chains of length 8-22 carbons.

3. The device of claim 1, wherein said hydrophilic polymer chains are polyethyleneglycol chains.