Arrays of proteins and methods of use thereof

• US 6,630,358 B1
• Filed: 05/12/2000
• Issued: 10/07/2003
• 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:
  US09/570,363
• Time in Patent Office: 1,243 Days
• Field of Search: 436/518, 436/524-535, 436/501, 435/4, 435/7.1, 422/131, 422/134
• US Class Current: 506/9
• 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 for the parallel, in vitro screening of biomolecular activity are provided. Methods of using the protein arrays are also disclosed. On the arrays, a plurality of different proteins, such as different members of a single protein family, are immobilized on one or more organic thinfilms on the substrate surface. The protein arrays are particularly useful in drug development, proteomics, and clinical diagnostics.

Claims

1. A method for screening a plurality of proteins for interactions with one or more components of a sample comprising the steps of:

providing a substrate having a surface, an array of discrete array-regions over said surface, an ordered hydrophobic-monolayer formed of alkyl-chains having alkyl-chain proximal ends which are chemisorbed or physisorbed to said surface, and opposite alkyl-chain distal ends, a hydrophilic-monolayer formed from hydrophilic-chains, each having a hydrophilic-chain proximal end by which each hydrophilic-chain is covalently linked to an alkyl-chain distal end, and an opposite hydrophilic-chain distal end, said ordered hydrophobic-monolayer and said hydrophilic-monolayer being effective in combination to resist non-specific protein binding, and, a plurality of protein-immobilizing groups covalently attached to a selected fraction of said hydrophilic-chains'"'"' distal ends within said array-regions;

applying said selected proteins to said protein immobilization regions 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 border regions resistant to non-specific protein binding to form a protein array resistant to non-specific protein binding;

exposing said protein regions to said fluid sample; and

,

detecting said interaction(s), if any, within said protein regions.

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

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