Skip to main content
SpringerLink
Log in

Purification and Autoactivation Method for Recombinant Coagulation Factor VII

  • Protocol
  • First Online:
Recombinant Glycoprotein Production

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1674))

Abstract

Recombinant coagulation factor VII is a very important and complex protein employed for treatment of hemophiliac patients (hemophilia A/B) who develop inhibitors antibodies to conventional treatments (FVIII and FIX). The rFVII is a glycosylated molecule and circulates in plasma as zymogen of 50 kDa. When activated the molecule is cleaved to 20–30 kDa and has a half-life of about 3 h, needing to be processed fast and efficiently until freeze-drying. Here, we describe a very simple and fast purification sequence for rFVII using affinity FVII Select resin and a dialysis system that can be easily scaled up.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Franchini M, Mannucci PM (2012) Past, present and future of hemophilia: a narrative review. Orphanet J Rare Dis 7:24

    Article  PubMed  PubMed Central  Google Scholar 

  2. Hedner U (2015) Recombinant activated factor VII: 30 years of research and innovation. Blood Rev Suppl 1:S4–S8

    Article  Google Scholar 

  3. Yoshihiro M, Takamiya O (2012) The expression of recombinant factor VII variants is influenced by different cells for transfection using expression studies. Rinsho Byori 60(3):193–200

    CAS  PubMed  Google Scholar 

  4. Eigenbrot C (2002) Structure, function, and activation of coagulation factor VII. Curr Protein Pept Sci 3(3):287–299

    Article  CAS  PubMed  Google Scholar 

  5. Bharadwaj D, Iino M, Kontoyianni M, Smith KJ, Foster DC, Kisiel W (1996) Factor VII central. A novel mutation in the catalytic domain that reduces tissue factor binding, impairs activation by factor Xa, and abolishes amidolytic and coagulant activity. J Biol Chem 271(48):30685–30691

    Article  CAS  PubMed  Google Scholar 

  6. Tiede A (2015) Half-life extended factor VIII for the treatment of hemophilia A. J Thromb Haemost Suppl 1:S176–S179

    Article  Google Scholar 

  7. Pedersen AH, Lund-Hansen T, Bisgaard-Frantzen H, Olsen F, Petersen LC (1989) Autoactivation of human recombinant coagulation factor VII. Biochemistry 28(24):9331–9336

    Article  CAS  PubMed  Google Scholar 

  8. Covas DT, Freitas MCC, Picanço-Castro V, Swiech K (2015) Processo de Produção de Fator VII de coagulação sanguínea e fator VII de coagulação sanguínea. Instituto Nacional da Propriedade Industrial (INPI) BR1020150123345

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Center for Cell-based Therapy CTC, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, Brazil

    Vladimir Granovski, Marcela C. C. Freitas & Dimas T. Covas

  2. Regional Blood Center of Ribeirão Preto, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, Brazil

    Marcela C. C. Freitas, Mario Soares Abreu-Neto & Dimas T. Covas

Authors
  1. Vladimir Granovski
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Marcela C. C. Freitas
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mario Soares Abreu-Neto
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Dimas T. Covas
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Vladimir Granovski .

Editor information

Editors and Affiliations

  1. Regional Blood Center of Ribeirão Preto, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, Brazil

    Virgínia Picanço-Castro

  2. School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, Brazil

    Kamilla Swiech

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media LLC

About this protocol

Cite this protocol

Granovski, V., Freitas, M.C.C., Abreu-Neto, M.S., Covas, D.T. (2018). Purification and Autoactivation Method for Recombinant Coagulation Factor VII. In: Picanço-Castro, V., Swiech, K. (eds) Recombinant Glycoprotein Production. Methods in Molecular Biology, vol 1674. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7312-5_18

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7312-5_18

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7311-8

  • Online ISBN: 978-1-4939-7312-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation