PMID: 9447247Feb 3, 1998Paper

Biochemical and functional characterization of recombinant von Willebrand factor produced on a large scale

Cellular and Molecular Life Sciences : CMLS
B E FischerF Dorner

Abstract

Recombinant von Willebrand factor (r-vWF) was produced in serum-free medium on a large scale in recombinant Chinese hamster ovary cells and was purified from fermentation supernatant by a combination of anion exchange chromatography and heparin affinity chromatography. Heparin affinity chromatography yielded r-vWF polymers of different degrees of multimerization. r-vWF was analysed by qualitative and quantitative functional analysis. We could show that while binding of r-vWF to platelets did not depend on multimerization of the molecule, ristocetin-induced platelet aggregation, binding to collagen and binding to heparin correlated directly with the extent of multimerization. Binding of recombinant coagulation factor VIII (r-FVIII) to r-vWF was studied by real-time biospecific interaction analysis and surface plasmon technology. The data indicated that binding of r-FVIII did not depend on r-vWF multimerization. Real-time biospecific interaction analysis suggested a potential stoichiometry of 2 to 2.5 r-vWF subunits per r-FVIII molecule. Kinetic analysis of the r-vWF-r-FVIII interaction gave a binding rate constant of 3 x 10(6) M-1 s-1 and an association constant of 2.5 x 10(9) M-1. Reaction of r-vWF with carbohydrate-specific le...Continue Reading

Citations

Aug 14, 2001·Transfusion clinique et biologique : journal de la Société française de transfusion sanguine·K Mertens
Feb 24, 1999·Haemophilia : the Official Journal of the World Federation of Hemophilia·J SiekmannH P Schwarz
Feb 24, 1999·Haemophilia : the Official Journal of the World Federation of Hemophilia·H P SchwarzP L Turecek
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