EPITOPE MAPPING OF ANTIBODIES TO VON WILLEBRAND FACTOR

两名von Willebrand因子抗体阳性患者的血浆 正在研究确定von Willebrand因子的表位，这些表位是 被抗体识别。一般的方法是 用酶法切割VWF，然后免疫沉淀 使用患者免疫球蛋白共价偶联到蛋白A的片段- 琼脂糖珠。凝血酶消化VWF后，大量 产生大小从约176kD到40kD的片段。 使用毛驴多克隆抗体的初步实验还没有 成功地免疫沉淀了任何VWF碎片。目前还不清楚 为什么，但可能的解释包括 消化和蛋白质不足，无法通过免疫印迹检测。 来自两名具有VWF抗体的患者的血浆将用于这些 研究什么时候VWF片段可以 用这种方法进行免疫沉淀。

Plasma from two patients with antibodies to von Willebrand factor are being studied to identify the epitopes on von Willebrand factor that are recognized by the antibodies. The general approach has been to enzymatically cleave vWf and then immunoprecipitate the resulting fragments using patient immunoglobulin covalently coupled to protein A - Sepharose beads. Following digestion of vWf by thrombin, numerous fragments ranging in size from about 176 kD to 40 kD are generated. Preliminary experiments using a burro polyclonal antibody have not been sucessful in immunoprecipitating any vWf fragments. It is unclear as to why, but possible explanations include destruction of the epitopes by digestion and insufficient protein to be detected by immunoblotting. Plasma from two patients with antibodies to vWf will be used in these studies when it is clear that the vWf fragments can be immunoprecipitated by this method.