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Quantification of Coagulation Factor VIII by Selective Reaction Monitoring

  • Edson Galdino do Nascimento Filho
  • Guilherme Pauperio Lanfredi
  • Mário Soares Abreu-Neto
  • Dimas Tadeu Covas
  • Vitor Marcel Faça
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1674)

Abstract

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 words

Mass spectrometry Selective reaction monitoring (SRM) Coagulation factor VIII Hemophilia A 

Notes

Acknowledgments

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.

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Copyright information

© Springer Science+Business Media LLC 2018

Authors and Affiliations

  • Edson Galdino do Nascimento Filho
    • 1
  • Guilherme Pauperio Lanfredi
    • 1
  • Mário Soares Abreu-Neto
    • 2
  • Dimas Tadeu Covas
    • 2
  • Vitor Marcel Faça
    • 1
  1. 1.Department of Biochemistry and ImmunologyRibeirão Preto Medical School, University of São PauloRibeirão PretoBrazil
  2. 2.Regional Blood Center of Ribeirão PretoRibeirão Preto Medical School, University of São PauloRibeirão PretoBrazil

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