The interaction of the factor VIII/von Willebrand factor complex (VIII/vWf), with guanidinium-derivatized matrices.
Five different guanidinium (Gu)-derivatized agarose matrices were investigated for their potential in chromatographically resolving the Factor VIII/von Willebrand complex, VIII/vWf, fibrinogen, Fg, and fibronectin, Fn, from cryoprecipitate. Using conventional NaCl gradient methodology it was found that the order of elution of specific plasma proteins, and the yield of VIII/vWf, varied with the methods used to derivatize the agarose beads. Good yields of VIII:C (generally 30-45%) were obtained with Gu-matrices prepared by bis-oxirane coupling procedures. Cryoprecipitate binding studies showed that the capacity of Gu-Sepharose 4B, prepared by isourea modification of amino-Sepharose 4B, was 36 units VIII/vWf per ml matrix. The product, depleted of both Fg and Fn, had a specific activity of 2 units VIII:C per mg total protein, (yield 100% vWf:Ag and 47% VIII:C).
Coagulation Research Laboratory, Rayne Institute, St. Thomas' Hospital, London, UK.
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Saundry
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Savidge
D002846:Chromatography, Affinity; D005169:Factor VIII / Q000302:isolation & purification*; D019791:Guanidine; D006146:Guanidines; D006863:Hydrogen-Ion Concentration; D012685:Sepharose; D014841:von Willebrand Factor / Q000302:isolation & purification*
D006146:Guanidines; D014841:von Willebrand Factor; D005169:Factor VIII; D012685:Sepharose; D019791:Guanidine