Methods of polypeptide identification, and compositions therefor

Patent metadata

Field	Value
Publication number	US-9880175-B2
Application number	US-201615390331-A
Country	US
Kind code	B2
Filing date	Dec 23, 2016
Priority date	Feb 13, 2006
Publication date	Jan 30, 2018
Grant date	Jan 30, 2018

Abstract

Methods are disclosed for identifying one or more proteins or polypeptides comprised by a sample. The methods comprise determining binding of each polypeptide with respect to each binding pool of a plurality of binding pools, wherein each binding pool comprises one or more probes which bind a structure comprised by a protein or polypeptide. In some aspects, polypeptides can be denatured and separated into individual polypeptide strands and immobilized on a solid support prior to determining binding of the binding pools. A protein, polypeptide or polypeptide strand can be identified by searching, in at least one database, for a protein or polypeptide sequence comprising binding pool targets either identical to or most similar to the binding pool targets comprised by the protein, polypeptide or polypeptide strand to be identified. Kits for identifying proteins, polypeptides and polypeptide strands are also disclosed.

First claim

What is claimed is:

1. A kit for identifying one or more polypeptides or polypeptide strands in a sample, the kit comprising:
a plurality of binding pools, wherein each binding pool comprises a plurality of probes, wherein each probe binds a binding pool target comprised by a protein or polypeptide described in a database comprising digital signatures for the polypeptides or polypeptide strands of the sample; and
a solid support, configured for immobilization of a plurality of protein or polypeptide strands comprised by a sample, whereby the protein or polypeptide strands immobilized on the solid support are separated by an optically resolvable distance from other protein or polypeptide strands comprised by the sample.

2. The kit of claim 1 , wherein a binding target of a probe comprised by a binding pool comprises a structure selected from the group consisting of a peptide motif comprising at least one amino acid up to about six amino acids, a glycosyl moiety of a glycopeptide or glycoprotein, a GPI anchor, a disulfide linkage, a pyroglutamic acid, a nitrotyrosine and any combination thereof, and wherein each probe is selected from the group consisting of at least one antibody, at a least one aptamer, at least one kinase, at least one avimer and any combination thereof.

3. The kit of claim 2 , wherein at least one probe of each binding pool comprises a label.

4. The kit of claim 3 , wherein the label is selected from the group consisting of a radioisotope, a chromophore, a fluorophore, a quantum dot, an enzyme and a resonance light scattering (RLS) particle.

5. The kit of claim 1 , comprising at least 5 binding pools.

6. The kit of claim 1 , comprising at least 10 binding pools.

7. The kit of claim 1 , comprising at least 12 binding pools.

8. The kit of claim 1 , comprising at least 15 binding pools.

9. The kit of claim 1 , further comprising at least one reducing agent.

10. The kit of claim 1 , further comprising at least one denaturant.

11. The kit of claim 1 , further comprising at least one secondary probe.

12. The kit of claim 11 , wherein the at least one secondary probe is a secondary antibody.

13. The kit of claim 12 , wherein the secondary antibody comprises a label.

14. The kit of claim 13 , wherein the label is a fluorophore.

15. The kit of claim 11 , wherein the at least one secondary probe is a resonance light scattering (RLS) particle.

16. The kit of claim 1 , wherein a binding pool recognizes multiple structures among the proteins or polypeptides of the sample or database.

17. The kit of claim 1 , wherein a binding pool recognizes approximately 50% of the proteins or polypeptides comprised by the database.

18. The kit of claim 1 , wherein the solid support is a glass microscope slide.

19. The kit of claim 1 , wherein the solid support is a plastic microscope slide.

20. The kit of claim 1 , wherein the solid support is an ELISA plate.

21. The kit of claim 1 , wherein the solid support is a microarray.

Assignees

• Univ Washington

Inventors

• Mitra Robi D

Classifications

• G01N33/6845 Primary

  Methods of identifying protein-protein interactions in protein mixtures · CPC title

• G01N33/54306

  Solid-phase reaction mechanisms · CPC title

• G01N33/582

  with fluorescent label · CPC title