Claims for Patent: 9,459,263

Summary for Patent: 9,459,263

Title:	VEGF-A.sub.121 assay
Abstract:	The invention provides a method for enriching the level of VEGF-A.sub.121 isoform in a sample by selectively removing the VEGF-A.sub.165 isoform from the sample using a neuropilin-1 pull-down procedure, then determining the total amount of VEGF-A remaining afterward. The invention provides methods of treating a patient suffering from a disease which may benefit from the administration of a VEGF antagonist by determining the level or ratio of VEGF-A.sub.121 in the patient\'s circulation. Methods of diagnosis, prognosis, monitoring, and patient stratification are also provided.
Inventor(s):	Papadopoulos; Nicholas (LaGrangeville, NY), Dore; Anthony (Tarrytown, NY), MacDonald; Douglas (New York, NY)
Assignee:	Regeneron Pharmaceuticals, Inc. (Tarrytown, NY)
Application Number:	14/811,056
Patent Claims:	1. A method of treating cancer, comprising, (a) obtaining a blood sample from a patient who suffers from cancer; (b) enriching the amount of VEGF-A121 in the sample by the steps of: contacting the sample comprising VEGF-A165 and VEGF-A121 with (1) a neuropilin 1 ("NRP1") protein, or (2) a fraction of the NRP1 protein comprising the B1B2 domain, wherein the VEGF-A165 binds to the NRP1 protein or fraction thereof to form a NRP1-VEGF-A165 complex; and separating the sample from the NRP1-VEGF-A165 complex to form a VEGF-A165-depleted sample; (c) quantifying the total remaining VEGF-A in the VEGF-A165-depleted sample; wherein the total remaining VEGF-A is enriched for VEGF-A121; and (d) administering a VEGF antagonist, if the total remaining VEGF-A is above a threshold amount. 2. The method of claim 1, wherein the NRP1 protein or fraction thereof is fixed to a substrate prior to step (b)(i). 3. The method of claim 2, wherein the substrate is a bead, a plate or a well of a microtiter plate. 4. The method of claim 3, wherein the bead, plate or well is coated with a solution of NRP1 protein. 5. The method of claim 4, wherein the bead, plate or well is coated with NRP1 protein by applying a 1 .mu.g/mL-10 .mu.g/mL solution of NRP1 protein to the plate or well. 6. The method of claim 1, wherein the NRP1 protein is human NRP1. 7. The method of claim 1, wherein the NRP1 protein is a fusion protein comprising a NRP1 protein domain comprising a B1B2 domain and an Fc domain. 8. The method of claim 7, wherein the NRP1-fusion protein consists essentially of NRP1 domains B1 and B2 fused to an Fc domain. 9. The method of claim 7, wherein the NRP1-fusion protein consists essentially of the extracellular domain of NRP1 fused to an Fc domain. 10. The method of claim 1, wherein the sample is additionally contacted with heparin at step (a). 11. The method of claim 10, wherein the heparin is at a concentration of about 1-10 .mu.g/mL. 12. The method of claim 1, wherein the VEGF-A165-depleted sample contains less than 65% of the VEGF-A165 of the non-depleted sample. 13. The method of claim 1, wherein the VEGF-A165-depleted sample contains less than 34% of the VEGF-A165 of the non-depleted sample. 14. The method of claim 1, wherein the blood sample comprises plasma or serum. 15. The method of claim 1, wherein the VEGF-A121-enriched fraction is quantified via ELISA using an anti-VEGF antibody. 16. The method of claim 1, wherein the VEGF antagonist is selected from the group consisting of bevacizumab, ranibizumab, aflibercept, and ramucirumab. 17. The method of claim 1, wherein the cancer is selected from the group consisting of metastatic breast cancer, gastric cancer, pancreatic cancer, renal cell carcinoma, non-small cell lung cancer, androgen-independent prostate cancer, ovarian cancer, adenocarcinoma, and colorectal cancer.

Details for Patent 9,459,263

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Genentech, Inc.	AVASTIN	bevacizumab	Injection	125085	02/26/2004		2033-05-22
Genentech, Inc.	LUCENTIS	ranibizumab	Injection	125156	06/30/2006		2033-05-22
Genentech, Inc.	LUCENTIS	ranibizumab	Injection	125156	08/10/2012		2033-05-22
Genentech, Inc.	LUCENTIS	ranibizumab	Injection	125156	10/13/2016		2033-05-22
Genentech, Inc.	LUCENTIS	ranibizumab	Injection	125156	03/20/2018		2033-05-22
Regeneron Pharmaceuticals, Inc.	EYLEA	aflibercept	Injection	125387	11/18/2011		2033-05-22
Regeneron Pharmaceuticals, Inc.	EYLEA	aflibercept	Injection	125387	08/16/2018		2033-05-22