Multiplex Array Useful for Assaying Protein-Protein Interaction

• US 20080274913A1
• Filed: 05/30/2006
• Published: 11/06/2008
• Est. Priority Date: 05/27/2005
• Status: Abandoned Application

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Invitrogen Corp. (Thermo Fisher Scientific Incorporated)
• Inventors: Lee, Kevin
• Application Number:
  US11/915,689
• Publication Number:
  US 20080274913A1
• Time in Patent Office: Days
• US Class Current: 506/14
• CPC Class Codes: G01N 33/5023 on expression patterns

Abstract

The described invention shows how multiple interactions between two proteins of interest can be determined by observing activation or lack thereof of intracellular proteins, following interaction of ligand and receptor.

Claims

1. A multiplex array useful in determining activity of a plurality of test proteins comprising:

a solid substrate having a plurality of receptacles, each of which contains a sample of cells, transformed or transfected with

(a) a first nucleic acid molecule comprising;

(i) a nucleotide sequence which encodes a first test protein,

(ii) a nucleotide sequence encoding a cleavage site for a protease, and

(iii) a nucleotide sequence which encodes a protein which activates a reporter gene in said cell,

(b) a second nucleic acid molecule which comprises;

(i) a nucleotide sequence which encodes a second test protein whose interaction with said first test protein in the presence of said test compound of interest is to be measured,

(ii) a nucleotide sequence which encodes a protease or a protease which is specific for said cleavage site,

wherein said first test protein differs from other first test proteins in each of said plurality of samples, wherein activity of said reporter gene is used to determine activity of one or more of said test proteins.

2. The multiplex array of claim 1, wherein each of said first test protein is a receptor.

3. The multiplex array of claim 2, wherein at least one of said first test proteins is a GPCR.

4. The multiplex array of claim 2, wherein each of said first test proteins is a GPCR.

5. The multiplex array of claim 2 where each of said first test proteins is a GPCR and each of said second test proteins is an arrestin.

6. The multiplex array of claim 1, comprising from about 25 to about 200 samples of cells, each of which is transformed or transfected with a different first test protein.

7. The multiplex array of claim 6, comprising from about 50 to about 200 samples of cells.

8. The multiplex array of claim 7, comprising from about 50 to about 100 samples of cells.

9. The multiplex array of claim 1, where at least a plurality of said first test proteins are selected from Table 1.

10. The multiplex array of claim 9, wherein at least a plurality of said first test proteins are selected from Table 2.

11. The multiplex array of claim 10, wherein said multiplex array comprises a plurality of test samples which express all of the first test proteins of Table 2.