Abstract
Protein C (PC) deficiency can cause thrombosis, inhibiting oxygen transport to tissue thus resulting in many complications, including death. Present treatment can cause catastrophic bleeding and other major medical problems. PC treatment has no bleeding or skin necrosis problems because it circulates in the blood as a zymogen and is only activated when and where it is needed.
The vitamin K dependent (VKD) proteins are homologous proteins, making the separation of PC from plasma extremely difficult. Immobilized metal affinity chromatography (IMAC) is investigated to separate the VKD proteins to replace immunoaffinity chromatography, because of the high cost of monoclonal antibodies.
An IDA-Cu column was found effective for the separation of PC from prothrombin, the most harmful contaminant. For Cohn fraction IV-1 separation, a DEAE column was found an efficient initial step, with about 25-fold PC purity increase. Following this step, an IDA-Cu column could remove many contaminants including prothrombin. The combination of DEAE and IDA-Cu resulted in PC purity increase of about 100-fold.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Esmon CT. The roles of protein C and thrombomodulin in the regulation of blood coagulation. J Bio Chem 1989;264:4743.
Bruley DF, Drohan WN. Protein C and related anticoagulants. Advances in applied biotechnology series, Volume 11, Gulf Publishing Company (1990).
Velander WH, Orthner CL, Tharakan JP, Madurawe RD, Ralston AH, Strickland DK, Drohan WN. Process implications for metal-dependent immunoaffinity interactions. Biotechnol Prog 1989;5:119–125.
Mantovaara T, Pertoft H, Porath J. Purification of factor VIII:c coagulant activity from rat liver nonparenchymal cell culture medium by immobilized metal ion affinity chromatography. Biotechnol Appl Biochem 1991;13:120–126.
Wu H, Bruley DF. Homologous human blood protein separation using immobilized metal affinity chromatography-protein C separation from prothrombin with application to the separation of factor IX and prothrombin. Biotechnol Prog 1999;15:928–931.
Wu H, Bruley DF, Kang KA. Protein C separation from human blood plasma Cohn fraction IV-1 using immobilized metal affinity chromatography. In:Hudetz AG, Bruley DF, editors. Advances in Experimental Medicine and Biology. Plenum:New York and London, 1998;454:697–704.
Tadepalli SS, Bruley DF, Kang KA, Drohan WN. Immobilized metal affinity chromatography process identification and scale-up for protein C production. In:Harrison DK, Delpy DT, editors. Advances in Experimental Medicine and Biology Plenum:New York and London, 1997;428:31–43.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2003 Springer Science+Business Media New York
About this chapter
Cite this chapter
Wu, H., Bruley, D.F. (2003). Protein C Separation from Human Blood Plasma Derivatives Using Low Cost Chromatography. In: Dunn, J.F., Swartz, H.M. (eds) Oxygen Transport to Tissue XXIV. Advances in Experimental Medicine and Biology, vol 530. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-0075-9_14
Download citation
DOI: https://doi.org/10.1007/978-1-4615-0075-9_14
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4613-4912-9
Online ISBN: 978-1-4615-0075-9
eBook Packages: Springer Book Archive