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Characterization and Novel Purification of Recombinant Human Protein C from Three Mammalian Cell Lines

Bio/Technology volume 8, pages 655–661 (1990)Cite this article

Abstract

Human Protein C (HPC), an antithrombotic factor with potential clinical utility, is a vitamin K-dependent protein that has several complex post-translational modifications. In an effort to define the functional roles of these modifications, recombinant HPC (rHPC) was expressed in and characterized from 3 adenovirus-transformed cell lines. The rHPC in crude culture medium from the 3 cell lines displayed anticoagulant activities that were either higher, slightly lower or much lower than that of plasma HPC. The rHPC from each cell line was purified and characterized using a novel, but simple chromatographic method, termed “pseudo-affinity”, capable of resolving molecules differing by only very slight modifications. We demonstrate the critical dependence of full γ-carboxylation on the function of this protein. In addition, our data indicate that both the γ-carboxyglutamate and glycosyl contents affect the functional activities of rHPC.

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Author notes

  1. S.C. Betty Yan: Corresponding author.

Authors and Affiliations

  1. Departments of Biochemistry and Molecular Genetics, Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN, 46285

    S.C. Betty Yan, Pam Razzano, Y. Bernice Chao, Jenna D. Walls, David T. Berg, Don B. McClure & Brain W. Grinnell

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  1. S.C. Betty Yan
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  2. Pam Razzano
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  3. Y. Bernice Chao
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  4. Jenna D. Walls
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  5. David T. Berg
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  7. Brain W. Grinnell
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Yan, S., Razzano, P., Chao, Y. et al. Characterization and Novel Purification of Recombinant Human Protein C from Three Mammalian Cell Lines. Nat Biotechnol 8, 655–661 (1990). https://doi.org/10.1038/nbt0790-655

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