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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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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DOI: https://doi.org/10.1038/nbt0790-655
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