Laboratory Testing for von Willebrand Factor Activity by Glycoprotein Ib Binding Assays (VWF:GPIb).

In addition to assessment of von Willebrand factor (VWF) antigen (VWF:Ag), the first-line laboratory investigation of possible von Willebrand disease (VWD) often includes an assay to measure GPIb (glycoprotein Ib) binding activity of VWF. A decreased GPIb binding activity is characteristic for most of the VWD types. For many years, the most frequently used assay for measuring GPIb binding activity was the ristocetin cofactor assay (VWF:RCo), which measures the agglutination of fixed human platelets by VWF in the presence of ristocetin. Because of performance issues, including high assay variability and a lack of VWF sensitivity, this assay is currently being replaced or supplemented by assays based on the binding of VWF to recombinant GPIb. One published method (now abbreviated VWF:GPIbR) uses wild-type GPIb for triggering the binding reaction in the presence of ristocetin. Another more widely used method (now abbreviated VWF:GPIbM) uses gain-of-function GPIb without ristocetin; this permits spontaneous binding of VWF to GPIb and avoids problems associated with the nonphysiological substance ristocetin. The binding of VWF to GPIb can be quantified by using different principles, e.g., ELISA, particle agglutination, or chemiluminescence. The following chapter describes a ristocetin-free method based on particle agglutination in more detail.