Mazurier C, Poulle M, Samor B, Hilbert L, Chtourou S
Laboratoire français du Fractionnement et des Biotechnologies, Lille and Les Ulis, France.
Vox Sang. 2004 Feb;86(2):100-4. doi: 10.1111/j.0042-9007.2004.00398.x.
Patients suffering from von Willebrand disease who are not responsive to desmopressin require substitutive treatment. This study was part of the development of a second-generation plasma-derived von Willebrand factor (VWF) concentrate, the manufacturing process of which includes two complementary viral-inactivation/elimination steps that are effective against non-enveloped viruses.
VWF was purified from solvent/detergent-treated cryoprecipitate through a combination of anion-exchange and affinity chromatography. The VWF preparation was diluted and filtered through filters with pore size of 35 nm. After concentration and formulation, the product was freeze-dried and further heated at 80 degrees C for 72 h. Tests were performed to evaluate the effects of nanofiltration and dry heating on VWF multimeric structure and function.
Nanofiltration and dry heating of the formulated product increased viral safety but did not modify VWF multimeric structure. Furthermore, these steps did not alter the ability of VWF to bind to platelet glycoprotein Ib, collagen and Factor VIII.
We have perfected a large-scale manufacturing process to produce a human plasma-derived VWF concentrate that boasts high specific activity and is very safe for the treatment of patients with von Willebrand disease.
对去氨加压素无反应的血管性血友病患者需要替代治疗。本研究是第二代血浆源性血管性血友病因子（VWF）浓缩物研发工作的一部分，其生产工艺包括两个互补的病毒灭活/去除步骤，对非包膜病毒有效。
通过阴离子交换和亲和层析相结合的方法，从经溶剂/去污剂处理的冷沉淀中纯化VWF。将VWF制剂稀释并通过孔径为35nm的滤器过滤。浓缩和制剂化后，产品冻干并在80℃进一步加热72小时。进行试验以评估纳滤和干热对VWF多聚体结构和功能的影响。
制剂产品的纳滤和干热提高了病毒安全性，但未改变VWF多聚体结构。此外，这些步骤未改变VWF与血小板糖蛋白Ib、胶原蛋白和因子VIII结合的能力。
我们完善了一种大规模生产工艺，以生产一种高比活性且治疗血管性血友病患者非常安全的人血浆源性VWF浓缩物。
