Capture arrays using polypeptide capture agents

• US 20060035277A1
• Filed: 07/31/2003
• Published: 02/16/2006
• Est. Priority Date: 02/01/2001
• Status: Abandoned Application

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Agilent Technologies, Inc.
• Inventors: Myerson, Joel
• Application Number:
  US10/632,870
• Publication Number:
  US 20060035277A1
• Time in Patent Office: Days
• US Class Current: 435/7.100
• CPC Class Codes: B01J 2219/00497 Features relating to the solid phase supports
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00574 radioactive
• CPC Class Codes: B01J 2219/00576 fluorophore
• CPC Class Codes: B01J 2219/00585 Parallel 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 2219/00626 Covalent
• CPC Class Codes: B01J 2219/0063 Other, e.g. van der Waals forces, hydrogen bonding
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• 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: B01J 2219/00727 Glycopeptides
• CPC Class Codes: C40B 30/04 by measuring the ability to specifically bind a target molecule, e.g. antibody-antigen binding, receptor-ligand binding
• 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: G01N 33/58 involving labelled substances G01N33/53 takes precedence
• CPC Class Codes: G01N 33/6845 Methods of identifying protein-protein interactions in protein mixtures

Abstract

A method of evaluating for the presence of a polypeptide in an analyte, using an addressable array of capture agents linked to a substrate. The analyte is exposed to the array and a set of fixed capture agents, such that the target molecules will bind to the array by means of the capture agents. After the target molecule has bound to the capture agents, it is modified using a label. The label does not interact or mark the capture agent. Kits using such arrays are further provided.

Claims

1-26. -26. (canceled)

27. An array of polypeptide capture agents, wherein said polypeptide capture agents are linked to a substrate and are free of a particular amino acid.

28. The array of claim 27, wherein said polypeptide capture agents are chemically modified so that they are free of said particular amino acid

29. The array of claim 27, wherein said particular amino acid is non-reactive with a labeling agent.

30. The array of claim 27, wherein said particular amino acid is chosen from arginine, lysine, cysteine, histidine, aspartic acid or glutamic acid.

31. The array of claim 27, wherein said particular amino acid is lysine.

32. The array of claim 27, wherein said polypeptide capture agents each contains a different amino acid sequence.

33. The array of claim 27, wherein said array comprises 10-100,000 polypeptide capture agents.

36. A system for evaluating the presence of a target polypeptide in a sample, said system comprising:

a) an array according to claim 27;

and

b) a labeling agent that does not covalently bind to said polypeptide capture agents of said array.

37. The system of claim 36, wherein said labeling agent covalently bonds to a reactive side chain of said particular amino acid.

38. The system of claim 37, wherein said particular amino acid is chosen from arginine, lysine, cysteine, histidine, aspartic acid or glutaminic acid.

39. The system of claim 38, wherein said particular amino acid is lysine.

40. The system of claim 36, wherein said labeling agent comprises a fluorophore.

41. The system of claim 36, wherein said labeling agent comprises a radioisotope.

42. The system of claim 36, wherein said system further comprises a target molecule that binds to a capture agent on said array.

43. The system of claim 42, wherein said labeling reagent binds to said target and does not bind to said capture agent.

34. An array of polypeptide capture agents, wherein said polypeptide capture agents are linked to a substrate and are made up of amino acids chosen from a group of amino acids that is free of arginine, lysine, cysteine, histidine, aspartic acid or glutamic acid.

35. An array of polypeptide capture agents, wherein said polypeptide capture agents are linked to a substrate and made up of amino acids chosen from:

ala, cys, asp, glu, phe, gly, his, ile, lys, leu, met, asp, pro, gln, ser, thr, val, trp, tyr;

ala, cys, asp, glu, phe, gly, his, ile, leu, met, asp, pro, gln, arg, ser, thr, val, trp, tyr;

ala, asp, glu, phe, gly, his, ile, lys, leu, met, asp, pro, gln, arg, ser, thr, val, trp, tyr;

ala, cys, asp, glu, phe, gly, ile, lys, leu, met, asp, pro, gln, arg, ser, thr, val, trp, tyr;

ala, cys, glu, phe, gly, his, ile, lys, leu, met, asp, pro, gln, arg, ser, thr, val, trp, tyr;

or

ala, cys, asp, phe, gly, his, ile, lys, leu, met, asp, pro, gln, arg, ser, thr, val, trp, tyr.