Arrays of protein-capture agents and methods of use thereof

• US 6,365,418 B1
• Filed: 05/18/2000
• Issued: 04/02/2002
• 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:
  US09/574,748
• Time in Patent Office: 684 Days
• Field of Search: 435/287.9, 435/286.1, 435/287.1, 435/288.3, 435/288.4, 435/7.1, 435/518, 435/524, 435/525, 435/527, 435/528, 435/532, 435/533, 435/535, 435/536, 427/261, 427/287, 427/387, 427/407.2, 427/134, 427/518, 427/487.2
• US Class Current: 506/18
• 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

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. An array device comprising

a substrate having a surface;

and,

one or more protein immobilization regions on said surface, said protein immobilization 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 immobilization regions, and opposite hydrophobic distal ends,

(ii) a hydrophilic monolayer attached to said ordered 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, and,

(iii) a plurality of functional groups attached to said hydrophilic/hydrophobic monolayers for covalently attaching protein-capture agents to said hydrophilic/hydrophobic monolayers within said immobilization regions;

wherein said hydrophobic monolayer and hydrophilic monolayers together are effective to resist non-specific protein binding.

2. The device of claim 1 wherein at least two of said protein immobilization regions are adjacent each other.

3. The device of claim 2 wherein at least two of said protein immobilization regions are separated from each other by one or more border regions.

4. The device of claim 3 wherein said border regions are formed from

(i) an ordered hydrophobic monolayer formed of alkyl chains having proximal ends which are chemisorbed or physisorbed to said surface within said immobilization regions, and opposite hydrophobic distal ends, and,

(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 making said border regions resistant to non-specific protein binding.

5. The device of claim 3 wherein said borders are hydrophobic.

6. The device of claim 3 wherein said borders are hydrophilic.