Methods for detecting receptor complexes comprising PI3K

• US RE44,437 E1
• Filed: 01/18/2012
• Issued: 08/13/2013
• Est. Priority Date: 04/01/2003
• Status: Expired due to Fees

• Current Assignee: Monogram Biosciences, Inc. (Laboratory Corporation Of America Holdings)
• Original Assignee: Monogram Biosciences, Inc. (Laboratory Corporation Of America Holdings)
• Inventors: Chan-Hui, Po-Ying, Dua, Rajiv, Mukherjee, Ali, Pidaparthi, Sailaja, Singh, Sharat, Salimi-Moosavi, Hossein, Shi, Yining
• Primary Examiner(s): Halvorson, Mark
• Application Number:
  US13/352,705
• Time in Patent Office: 573 Days
• Field of Search: None
• US Class Current: 435/7.23
• CPC Class Codes: G01N 2333/485 Epidermal growth factor [EGF] (urogastrone)
• CPC Class Codes: G01N 2333/49 Platelet-derived growth factor [PDGF]
• CPC Class Codes: G01N 2800/52 Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
• CPC Class Codes: G01N 33/542 with steric inhibition or signal modification, e.g. fluorescent quenching
• CPC Class Codes: G01N 33/57492 involving compounds localized on the membrane of tumor or cancer cells
• CPC Class Codes: G01N 33/6803 General methods of protein analysis not limited to specific proteins or families of proteins
• CPC Class Codes: G01N 33/6842 Proteomic analysis of subsets of protein mixtures with reduced complexity, e.g. membrane proteins, phosphoproteins, organelle proteins
• CPC Class Codes: G01N 33/6872 Intracellular protein regulatory factors and their receptors, e.g. including ion channels

Abstract

The invention is directed to a new class of biomarker in patient samples comprising dimers of ErbB cell surface membrane receptors. In one aspect, the invention includes a method of determining the status of a disease or healthful condition by correlating such condition to amounts of one or more dimers of ErbB cell surface membrane receptors measured directly in a patient sample, in particular a fixed tissue sample. In another aspect, the invention includes a method of determining a status of a cancer in a specimen from an individual by correlating measurements of amounts of one or more dimers of ErbB cell surface membrane receptors in cells of the specimen to such status, including presence or absence of a pre-cancerous state, presence or absence of a cancerous state, prognosis of a cancer, or responsiveness to treatment. Preferably, methods of the invention are implemented by using sets of binding compounds having releasable molecular tags that are specific for multiple components of one or more types of receptor dimers. After binding, molecular tags are released and separated from the assay mixture for analysis.

Claims

1. A method for detecting in a biological sample the presence or amount of a receptor complex comprising a phosphatidylinositol-3-kinase (PI3K) protein and a receptor, comprising:

a. contacting with the sample a binding compound having a molecular tag attached thereto by a cleavable linkage and a cleaving probe having a cleavage inducing-moiety with an effective proximity, wherein the binding compound specifically binds either the PI3K or the receptor and the cleaving probe specifically binds an opposite member of the receptor complex from that bound by the cleaving probe;

b. cleaving the cleavable linker of the binding compound within the effective proximity of the cleavage-inducing moiety of the cleaving probe, thereby releasing the molecular tag; and

c. identifying the released molecular tag to determine the presence or amount of the receptor complex comprising the PI3K and the receptor in the sample.

2. The method of claim 1, wherein the receptor is Her1.

3. The method of claim 1, wherein the binding compound comprises an anti-PI3K antibody.

4. The method of claim 1, wherein the binding compound comprises an anti-Her3 antibody.

5. The method of claim 1, wherein the cleaving probe comprises an anti-PI3K antibody.

6. The method of claim 1, wherein the cleaving probe comprises an anti-Her3 antibody.

7. The method of claim 1, wherein the biological sample comprises cancer cells.

8. The method of claim 7, wherein the cancer cells are obtained from a patient.

9. The method of claim 8, wherein the biological sample is obtained by a biopsy of the cancer.

10. The method of claim 8, wherein the biological sample is obtained by purifying circulating cancer cells from the patient'"'"'s blood.

11. The method of claim 7, wherein the cancer is a breast cancer.

12. The method of claim 1, wherein the step of detecting is preceded by electrophoretically separating the molecular tag.

13. The method of claim 1, wherein the cleavage-inducing moiety is a photosensitizer.

14. The method of claim 1, wherein the receptor is Her2.

15. The method of claim 1, wherein the receptor is Her3.

16. The method of claim 1, wherein the binding compound comprises an anti-Her1 antibody.

17. The method of claim 1, wherein the binding compound comprises an anti-Her2 antibody.

18. The method of claim 1, wherein the cleaving probe comprises an anti-Her1 antibody.

19. The method of claim 1, wherein the cleaving probe comprises an anti-Her2 antibody.

20. The method of claim 7, wherein the cancer is a lung cancer.