Chromatographic purification and properties of a therapeutic human protein C concentrate

M. Radosevich, F. L. Zhou, J. J. Huart, T. Burnouf

Research output: Contribution to journalArticlepeer-review

7 Citations (Scopus)


Protein C deficiency (inherited and acquired) has a relatively high incidence rate in the general population worldwide. For many years, protein C deficient patients have been treated with fresh frozen plasma, prothrombin complex concentrates, heparin or oral anticoagulants, which all have clinical drawbacks. We report the production process of a highly purified human protein C concentrate from 1500 l of cryo-poor plasma by a four-step chromatographic procedure. After DEAE-Sephadex adsorption, protein C was separated from clotting factors II, VII and IX by DEAE-Sepharose FF and further purified, using a new strategy, by an on-line chromatographic system combining DMAE-Fractogel and heparin-Sepharose CL-6B. In addition, the product was treated against viral risks by solvent-detergent and nanofiltration on 15-nm membranes. The protein C concentrate was essentially free of other vitamin K-dependent proteins. Proteolytic activity was undetectable. Neither activated protein C, prekallikrein activator, nor activated vitamin K-dependent clotting factors were found resulting in good stability of the protein C activity. In vitro and in vivo animal tests did not reveal any sign of potential thrombogenicity. The final freeze-dried product had a mean protein C concentration of 58 IU/ml and a mean specific activity of 215 IU/mg protein, corresponding to over 12 000-fold purification from plasma. Therefore, this concentrate appears to be of potential benefit for the treatment of protein C deficiency.

Original languageEnglish
Pages (from-to)199-207
Number of pages9
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Issue number1-2
Publication statusPublished - Jun 25 2003
Externally publishedYes


  • Factor II
  • Factor IX
  • Factor VII
  • Protein C
  • Proteins

ASJC Scopus subject areas

  • Biochemistry


Dive into the research topics of 'Chromatographic purification and properties of a therapeutic human protein C concentrate'. Together they form a unique fingerprint.

Cite this