Lombardi R, Gelfi C, Righetti P G, Lattuada A, Mannucci P M
Thromb Haemost. 1986 Apr 30;55(2):246-9.
A new electrophoretic method is described for rapid screening of abnormalities of the multimeric structure of von Willebrand factor in von Willebrand's disease. The method is based on the transfer of the separated proteins from agarose gels onto nitrocellulose foils followed by immunoperoxidase staining. It has the advantage of not requiring radio-iodinated antibodies and reduces the working time for the entire procedure from 5-6 days to 3 days. Electroblotting followed by immunoperoxidase staining differentiates patients with intact multimeric structure from those without intermediate and/or large multimers. The more subtle defects of the inner structure of the smallest multimers found in patients with type II von Willebrand's disease can also be identified. A potential disadvantage of electroblotting and immunoperoxidase staining is the lesser sensitivity of this technique, which results in the detection of a smaller number of multimers (11-12 bands) than by autoradiography without transfer onto nitrocellulose (16-17 bands).
本文描述了一种新的电泳方法，用于快速筛查血管性血友病患者血管性血友病因子多聚体结构异常。该方法基于将分离的蛋白质从琼脂糖凝胶转移到硝酸纤维素膜上，然后进行免疫过氧化物酶染色。它的优点是不需要放射性碘化抗体，并将整个过程的工作时间从5 - 6天缩短至3天。电转印后进行免疫过氧化物酶染色可区分多聚体结构完整的患者与没有中间和/或大多聚体的患者。在II型血管性血友病患者中发现的最小多聚体内层结构的更细微缺陷也能够被识别。电转印和免疫过氧化物酶染色的一个潜在缺点是该技术的灵敏度较低，与未转移到硝酸纤维素膜上的放射自显影法相比，检测到的多聚体数量较少（11 - 12条带），后者可检测到16 - 17条带。
