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Thromb Haemost 1986; 55(03): 318-324
DOI: 10.1055/s-0038-1661555
Original Article
Schattauer GmbH Stuttgart

A Two-Site, Monoclonal Antibody-Based Immunoassay for von Willebrand Factor -Demonstration that vWF Function Resides in a Conformational Epitope

S Chand
*   The Academic Department of Immunology, Royal Free Hospital School of Medicine, London, UK
,
A McCraw
+   Katherine Dormandy, Haemophilia Centre, Royal Free Hospital School of Medicine, London, UK
,
R Hutton
+   Katherine Dormandy, Haemophilia Centre, Royal Free Hospital School of Medicine, London, UK
,
E G D Tuddenham
+   Katherine Dormandy, Haemophilia Centre, Royal Free Hospital School of Medicine, London, UK
,
A H Goodall
*   The Academic Department of Immunology, Royal Free Hospital School of Medicine, London, UK
› Author Affiliations
Further Information

Publication History

Received 21 January 1986

Accepted after revision 06 March 1986

Publication Date:
26 July 2018 (online)

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Summary

Two monoclonal antibodies (RFF-VIII: R/l and RFF-VIII:R/ 2) which recognise the same epitope on von Willebrand factor (vWF) have been used in a simple, two-site, solid-phase immunoradiometric (IRMA) or enzyme-linked assay (ELISA) to analyse vWF in plasma from normal individuals and from patients with von Willebrand’s disease (vWD). Results obtained confirm our previous findings (using RFF-VIII :R/2 in a one-site, fluid-phase IRMA) that the MAbs detect the presence of an epitope on the vWF molecule that reflects its function. This epitope is involved in vWF binding to the GPIb protein on platelets. It is reduced in all types of vWD, including type II (or variant) vWD. It is present in normal plasma, in vWF released from normal platelets and from cultured umbilical cord vein endothelial cells. The epitope is, however, found to be reduced in serum. Studies on SDS-treated vWF prove that this GPIb-binding site is dependent on the conformation of the vWF multimers.

Keywords

von Willebrand factor - Monoclonal antibodies - Immunoassay
 

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