School of Medical Sciences, Faculty of Medicine and Health University of Sydney, Westmead Hospital, Westmead, NSW, Australia.
School of Dentistry and Medical Sciences, Faculty of Science and Health, Charles Sturt University, Wagga, Wagga, NSW, Australia.
Methods Mol Biol. 2023;2663:679-691. doi: 10.1007/978-1-0716-3175-1_45.
von Willebrand factor (VWF) is a large adhesive plasma protein that expresses several functional activities. One of these activities is to bind coagulation factor VIII (FVIII) and to protect it from degradation. Deficiency of, and/or defects in, VWF can give rise to a bleeding disorder called von Willebrand disease (VWD). The defect in VWF that affects its ability to bind to and protect FVIII is captured within type 2N VWD. In these patients, FVIII is produced normally; however, plasma FVIII quickly degrades as it is not bound to and protected by VWF. These patients phenotypically resemble those with hemophilia A, where instead, FVIII is produced in lower amount. Both hemophilia A and 2N VWD patients therefore present with reduced levels of plasma FVIII relative to VWF level. However, therapy differs, since patients with hemophilia A are given FVIII replacement products, or FVIII mimicking products; instead, patients with 2N VWD require VWF replacement therapy, since FVIII replacement will only be effective for a short term, given this replacement product will quickly degrade in the absence of functional VWF. Thus, 2N VWD needs to be differentiated from hemophilia A. This can be achieved by genetic testing or by use of a VWF:FVIII binding assay. The current chapter provides a protocol for the performance of a commercial VWF:FVIII binding assay.
血管性血友病因子(VWF)是一种大型黏附性血浆蛋白,具有多种功能活性。其中一种活性是结合凝血因子 VIII(FVIII)并保护其免受降解。VWF 的缺乏和/或缺陷可导致一种称为血管性血友病(VWD)的出血性疾病。影响 VWF 结合和保护 FVIII 能力的 VWF 缺陷存在于 2N 型 VWD 中。在这些患者中,FVIII 正常产生;然而,由于它未与 VWF 结合和保护,血浆 FVIII 迅速降解。这些患者在表型上类似于血友病 A 患者,而血友病 A 患者中 FVIII 的产生量较低。因此,与 VWF 水平相比,这两种疾病患者的血浆 FVIII 水平均降低。然而,治疗方法不同,因为血友病 A 患者给予 FVIII 替代产品或 FVIII 模拟产品;而 2N VWD 患者需要 VWF 替代治疗,因为在缺乏功能性 VWF 的情况下,FVIII 替代仅在短期内有效。因此,需要将 2N VWD 与血友病 A 区分开来。这可以通过基因检测或使用 VWF:FVIII 结合测定来实现。本章提供了一种商业 VWF:FVIII 结合测定的操作方案。