Structure and Function of a Recombinant von Willebrand Factor Drug Candidate

Peter L Turecek; Gerald Schrenk; Hanspeter Rottensteiner; Katalin Varadi; Edouard Bevers; Peter Lenting; Nicola Ilk; Uwe B Sleytr; Hartmut J Ehrlich; Hans Peter Schwarz

doi:10.1055/s-0030-1255445

Seminars in Thrombosis and Hemostasis, Inhaltsverzeichnis

Semin Thromb Hemost 2010; 36(5): 510-521
DOI: 10.1055/s-0030-1255445

Structure and Function of a Recombinant von Willebrand Factor Drug Candidate

Peter L. Turecek¹ , Gerald Schrenk¹ , Hanspeter Rottensteiner¹ , Katalin Varadi¹ , Edouard Bevers³ , Peter Lenting⁴ , Nicola Ilk² , Uwe B. Sleytr² , Hartmut J. Ehrlich¹ , Hans Peter Schwarz¹

¹Baxter BioScience, Vienna, Austria
²Department of NanoBiotechnology, BOKU-University of Natural Resources and Applied Life Sciences, Vienna, Austria
³Cardiovascular Research Institute Maastricht, Department of Biochemistry, Maastricht, The Netherlands
⁴INSERM U770, Le Kremlin-Bicêtre, France

Abstract

ABSTRACT

The complex structure, large size, and multiple posttranslational modifications of von Willebrand factor (VWF) presented a technological challenge for the production of recombinant VWF (rVWF). Nonetheless, we developed an rVWF product for treating von Willebrand disease, whereupon rVWF is coexpressed with recombinant factor VIII (rFVIII) in Chinese hamster ovary cells used to produce rFVIII for the treatment of hemophilia A. Here we describe the characterization of the structure and function of the rVWF drug product, with a focus on its in vitro platelet aggregation and matrix protein binding functions. Electron microscopy and multimer analysis revealed a highly organized structure for the rVWF protein, with a homogeneous multimer distribution including ultrahigh molecular weight multimers. The specific activity for binding to collagen and platelets mediated by ristocetin is higher in rVWF than in commercial plasma-derived VWF-FVIII complex products. The affinity and binding capacity of rVWF to FVIII is comparable to VWF in plasma. rVWF effectively binds to platelets and promotes platelet adhesion under shear stress similar to VWF in human plasma.

KEYWORDS

Recombinant von Willebrand factor - VWF - von Willebrand disease - VWD - hemophilia - structure - platelet binding

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Referenzen

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Peter L TurecekPh.D.

Baxter Innovations GmbH, Industriestrasse 67

1220, Vienna, Austria

eMail: peter_turecek@baxter.com