Large scale purification of factor X by hydrophobic chromatography.
Journal Article (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
International Standard Serial Number (ISSN)
- 0032-7484
Digital Object Identifier (DOI)
- 10.1080/00327488808062531
Language
- eng
Conference Location
- United States