Abstract
The major human vitamin K-dependent proteins were purified from plasma using immunoadsorbents made with antibodies specific for each protein. Monoclonal antibodies to Factor VII, Factor IX, Factor X, Protein C, and Protein S were prepared from mice immunized with isolated vitamin K-dependent antigens. Purified monoclonal antibodies and a purified burro polyclonal anti-prothrombin immunoglobulin were individually coupled to Sepharose and used in a tandem series of columns to purify each of the vitamin K-dependent proteins from eluates of barium citrate precipitates of plasma. The proteins were eluted from the columns by sodium thiocyanate and retained functional activity following dialysis. Prothrombin, Factor VII, Factor IX, Factor X and Protein C were essentially homogeneous as judged by NaDodSO4-PAGE; Protein S was isolated as a Protein S-C4b binding protein complex. These results indicate the utility of monoclonal antibody immunoadsorbents for purifying the human vitamin K-dependent proteins and represent a considerable simplification over other purification schemes.
References
Mar 6, 1979·Biochemistry·R G DiScipio, E W Davie
May 15, 1979·Biochemistry·J W Bloom, K G Mann
Sep 1, 1979·The Journal of Clinical Investigation·W Kisiel
Apr 1, 1976·European Journal of Immunology·G KöhlerC Milstein
Feb 22, 1977·Biochemistry·R G Di ScipioE W Davie
Feb 1, 1976·The British Journal of Radiology·G O'Reilly, K Jefferson
Jan 1, 1976·Methods in Enzymology·R L LundbladK G Mann
Jun 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·H A LiebmanB Furie
May 15, 1985·Thrombosis Research·W R Church, K G Mann
Mar 30, 1973·Biochimica Et Biophysica Acta·W Kisiel, D J Hanahan
Jul 1, 1974·Analytical Biochemistry·S C MarchP Cuatrecasas
Aug 15, 1970·Nature·U K Laemmli
Jan 1, 1983·Preparative Biochemistry·S P BajajS F Brown
Apr 1, 1981·Proceedings of the National Academy of Sciences of the United States of America·B Dahlbäck, J Stenflo
Feb 1, 1984·Analytical Biochemistry·L V Rao, S P Bajaj
Mar 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·C A Fulcher, T S Zimmerman
Dec 1, 1982·Thrombosis Research·W B FosterK G Mann
Jan 1, 1981·Proceedings of the National Academy of Sciences of the United States of America·J A KatzmannK G Mann
Jan 1, 1981·Preparative Biochemistry·S P BajajC Prodanos
Citations
Mar 1, 1996·Thrombosis Research·W T SumnerM Hoffman
Feb 7, 2004·Journal of Chromatography. a·Myles LindsayKevin E Van Cott
Aug 11, 1989·Journal of Chromatography·J X Huang, G Guiochon
Jun 8, 2001·Journal of Chromatography. B, Biomedical Sciences and Applications·H Husi, M D Walkinshaw
Feb 17, 2000·Journal of Chromatography. B, Biomedical Sciences and Applications·H Husi, M D Walkinshaw
Oct 1, 1994·British Journal of Haematology·D M MonroeM Hoffman
Jan 1, 1990·Vox Sanguinis·J TharakanD B Clark
Jun 18, 2010·Arteriosclerosis, Thrombosis, and Vascular Biology·Kenneth G Mann
Sep 7, 2012·Thrombosis Research·Matthew GisselSaulius Butenas
May 25, 2010·Journal of Thrombosis and Haemostasis : JTH·T OrfeoK G Mann
Jan 1, 1990·The American Journal of Medicine·W T Branch, A Suchman
Sep 15, 1991·Thrombosis Research·V RegnaultJ F Stoltz
Dec 24, 2013·Biochimica Et Biophysica Acta·Beth A BouchardSaulius Butenas
Nov 30, 2007·Academic Medicine : Journal of the Association of American Medical Colleges·Larry A GreenPerry A Pugno
Feb 25, 2011·Academic Medicine : Journal of the Association of American Medical Colleges·Nicholas Kontos
Apr 18, 1994·Annals of the New York Academy of Sciences·K G Mann