Chromatographic purification and properties of a therapeutic human protein C concentrate

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

研究成果: 雜誌貢獻文章

6 引文 斯高帕斯(Scopus)

摘要

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.
原文英語
頁(從 - 到)199-207
頁數9
期刊Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
790
發行號1-2
DOIs
出版狀態已發佈 - 六月 25 2003
對外發佈Yes

ASJC Scopus subject areas

  • Biochemistry

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