Quantification of Coagulation Factor VIII by Selective Reaction Monitoring
- 1.9k Downloads
Coagulation factor VIII (FVIII) is an important glycoprotein involved in the extrinsic coagulation cascade. Mutations in FVIII gene results in hemophilia A, a recessive coagulation disorder that is clinically managed by administration of purified FVIII from blood donors or recombinant FVIII. Because of its fundamental therapeutic application, biotechnological production of FVIII requires rigid quality control and monitoring in patients and clinical trials. Here, we describe a protocol for a mass spectrometry based approach termed selective reaction monitoring (SRM) as an important alternative tool for accurate and sensitive quantitation of purified or recombinant FVIII.
Key wordsMass spectrometry Selective reaction monitoring (SRM) Coagulation factor VIII Hemophilia A
This research was supported by FAPESP (Young Scientist Grants—Proc. No. 2011/09347-1), Center for Cell Based Thereapy—CTC-CEPID (Proc. FAPESP 2013/08135-2) and CISBi-NAP. G.P.L. received a fellowship from CNPq Proc. No. 130142/2015-8, and V.M.F. received a fellowship from CNPq, Proc. No. 308561/2014-7.
- 7.Russo-Carbolante EMS, Swiech K, Covas DT, Picanço-Castro V (2015) Large scale and stable production of human FVIII in the human cell line Sk-Hep-1. US Patent 8,969,041 B2Google Scholar