Claims for Patent: 10,577,641

Summary for Patent: 10,577,641

Title:	Truncated Her2 SRM/MRM assay
Abstract:	This disclosure provides ten (10) specific peptides, and particular peptide characteristics, from the cell membrane-bound Her2 protein and a diagnostic assay useful for determining the presence and amount of full length and truncated versions of the full-length Her2 protein in cells derived from formalin fixed paraffin embedded tissue.
Inventor(s):	Krizman; David (Gaithersburg, MD)
Assignee:	EXPRESSION PATHOLOGY, INC. (Rockville, MD)
Application Number:	15/707,283
Patent Claims:	1. A method for measuring the amount of truncated and full-length Her2 protein in a human biological sample of formalin-fixed tissue, comprising detecting and quantifying by mass spectrometry an amount of a first fragment peptide from the intracellular domain and a second fragment peptide from the extracellular domain of the Her2 protein in a protein digest prepared from said biological sample; and calculating the level of said first and said second fragment peptides in said sample, wherein said first fragment peptide is the peptide of SEQ ID NO:7 and said second peptide is selected from the group consisting of SEQ ID NOs: 1-5; wherein the level of said first and said second fragment peptides correlates to the amount of the truncated and full-length Her 2 protein in said biological sample. 2. The method of claim 1, further comprising the step of fractionating said protein digest prior to detecting said peptides. 3. The method of claim 2, wherein said fractionating step is selected from the group consisting of liquid chromatography, nano-reversed phase liquid chromatography, high performance liquid chromatography and reverse phase high performance liquid chromatography. 4. The method of claim 1, wherein said protein digest comprises a protease digest. 5. The method of claim 4, wherein said protein digest comprises a trypsin digest. 6. The method of claim 1, wherein said mass spectrometry comprises tandem mass spectrometry, ion trap mass spectrometry, triple quadrupole mass spectrometry, MALDI-TOF mass spectrometry, MALDI mass spectrometry, and/or time of flight mass spectrometry. 7. The method of claim 1, wherein the tissue is paraffin embedded tissue. 8. The method of claim 1, wherein the tissue is obtained from a tumor. 9. The method of claim 8, wherein the tumor is a primary tumor. 10. The method of claim 8, wherein the tumor is a secondary tumor. 11. The method of claim 1, wherein quantifying said first and said second Her2 fragment peptides comprises comparing the amount of said first and said second Her2 fragment peptides in the biological sample to the amount of the same Her2 fragment peptides in a different and separate biological sample. 12. The method of claim 1, wherein quantifying said first and said second Her2 fragment peptides comprises determining the amount of said Her2 fragment peptides in the biological sample by comparing to spiked internal standard peptides of known amount, wherein both the native peptides in the biological sample and the corresponding internal standard peptides have the same amino acid sequence. 13. The method of claim 12, wherein each internal standard peptide is an isotopically labeled peptide. 14. The method of claim 13, wherein each isotopically labeled internal standard peptide comprises one or more heavy stable isotopes selected from .sup.18O, .sup.17O, .sup.34S, .sup.15N, .sup.13C, .sup.2H and a combination thereof. 15. The method of claim 1, further comprising obtaining the biological sample from a subject, wherein detecting and quantifying said first and said second Her2 fragment peptides in the protein digest indicates the presence of the full-length Her2 protein, presence of truncated Her2 protein, and/or combination of the presence of both and an association with cancer in the subject. 16. The method of claim 15, further comprising correlating the detected and quantitated amounts of said Her2 fragment peptides to the diagnostic stage/grade/status of the cancer. 17. The method of claim 1, further comprising selecting a treatment for a subject based on the presence, absence, or quantified levels of said Her2 fragment peptides in the protein digest. 18. The method of claim 1, further comprising administering to a subject a therapeutically effective amount of a therapeutic agent targeted to the Her2 protein, wherein the treatment decision about which agent and amount of agent used for treatment is based upon the level of the full-length Her2 protein or truncated Her2 protein in the biological sample wherein said therapeutic agent is selected from the group consisting of trastuzumab and lapatinib. 19. The method of claim 1, wherein detecting and quantitating said first and said second Her2 fragment peptides can be combined with detecting and quantitating other peptides from other proteins in multiplex.

Details for Patent 10,577,641

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Genentech, Inc.	HERCEPTIN	trastuzumab	For Injection	103792	09/25/1998		2030-12-08
Genentech, Inc.	HERCEPTIN	trastuzumab	For Injection	103792	02/10/2017		2030-12-08