Large scale purification of factor X by hydrophobic chromatography.

Published

Journal Article

Factor X is a critical enzyme in the blood coagulation cascade, however, in recent years the coagulation zymogen factor X has received additional interest as a selective proteinase to allow production of functional eukaryotic proteins in a prokaryotic expression system. Traditional factor X purification schemes suffer from low yields, low capacity, lengthy dialysis steps, and contamination by the autoproteolytic activated enzyme factor Xa. By incorporating a reversible inhibitor of factor X activation, we were able to recover 67% of the factor X present without any detectable activated enzyme. Six liters of plasma could be processed onto a 50 mL phenylalanine-Sepharose hydrophobic chromatography column without saturating the matrix. The final product is devoid of detectable proteolytic activity. At time of use, the zymogen is specifically activated with a Sepharose-bound activating enzyme isolated from Russell's Viper Venom, resulting in factor Xa free of other detectable proteinases.

Full Text

Duke Authors

Cited Authors

  • Friedberg, RC; Pizzo, SV

Published Date

  • 1988

Published In

Volume / Issue

  • 18 / 3

Start / End Page

  • 303 - 320

PubMed ID

  • 3237647

Pubmed Central ID

  • 3237647

International Standard Serial Number (ISSN)

  • 0032-7484

Digital Object Identifier (DOI)

  • 10.1080/00327488808062531

Language

  • eng

Conference Location

  • United States