EPITOPE MAPPING OF ANTIBODIES TO VON WILLEBRAND FACTOR

来自两名具有血管性血友病因子抗体的患者的血浆， 正在研究以确定血管性血友病因子上的表位， 被抗体识别。 一般的做法是， 酶促裂解vWf，然后免疫沉淀所得的 使用共价偶联至蛋白A的患者免疫球蛋白的片段 琼脂糖珠。 在凝血酶消化vWf后， 产生大小在约176 kD至40 kD范围内的片段。 使用驴多克隆抗体的初步实验尚未完成。 成功免疫沉淀任何vWf片段。 目前还不清楚 为什么，但可能的解释包括表位的破坏， 消化和免疫印迹检测不到足够的蛋白质。 将在这些研究中使用来自两名具有vWf抗体的患者的血浆。 研究表明，当vWf片段可以被 通过这种方法免疫沉淀。

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.