Yoshioka A, Sakai T, Yamamoto K, Ohkubo Y, Fukui H
Department of Pediatrics, Nara Medical College, Japan.
Thromb Haemost. 1987 Aug 4;58(2):705-8.
A simple method for analyzing the activation mechanism of FIX in patients with hemophilia B variants is described. The procedure consists of rapid partial purification of FIX by BaCl2 adsorption-elution from only 3 ml of plasma, incubation with FXIa/Ca2+, SDS-PAGE, western blotting and subsequent autoradiography using monoclonal anti-FIX antibody. Abnormal FIX from the plasma of 7 unrelated patients with hemophilia BR, B+ or BM was investigated. A time course study showed that FIX in the patient with hemophilia BM (Nagoya I), BM (Nagoya II) and B Kawachinagano seemed not to be cleaved by FXIa, FIX in the patient with hemophilia B Kashihara was partially cleaved, FIX in the patient with hemophilia BM (Takatsuki) showed delayed cleavage, and that FIX in the patient with hemophilia BM (Niigata) and BM (Kiryu) was cleaved completely at a rate similar to normal FIX. These findings were identical to those previously observed for the respective factors in a purified system. The procedure used here is useful for screening for a defective activation mechanism of abnormal FIX.
描述了一种分析B型血友病变异患者FIX激活机制的简单方法。该程序包括仅从3毫升血浆中通过BaCl2吸附 - 洗脱快速部分纯化FIX，与FXIa/Ca2+孵育，SDS - PAGE，蛋白质印迹以及随后使用单克隆抗FIX抗体进行放射自显影。研究了7名无关的B型血友病BR、B+或BM患者血浆中的异常FIX。时间进程研究表明，BM型（名古屋I）、BM型（名古屋II）和川内野B型血友病患者的FIX似乎未被FXIa裂解，柏原B型血友病患者的FIX部分被裂解，高槻BM型血友病患者的FIX显示出延迟裂解，而新潟BM型和桐生BM型血友病患者的FIX以与正常FIX相似的速率完全裂解。这些发现与先前在纯化系统中对各自因子观察到的结果一致。这里使用的程序对于筛选异常FIX的缺陷激活机制很有用。
