Yoshioka A, Ohkubo Y, Nishimura T, Tanaka I, Fukui H, Ogata K, Kamiya T, Takahashi H
Thromb Res. 1986 Jun 1;42(5):595-604. doi: 10.1016/0049-3848(86)90338-5.
Abnormal factor IX was isolated from the plasma of a patient with hemophilia B Kashihara and two patients with hemophilia BM. The F.IX was purified to homogeneity by using monoclonal anti-F.IX-Sephrose, heparin-Sepharose and DEAE-Sephadex A-50 affinity chromatography successively. The isolated proteins have the same molecular weight and the same mobility on crossed immunoelectrophoresis as normal F.IX. The limited proteolysis of purified proteins was induced by F.XIa/Ca2+ or by RVV-X/Ca2+. A time course study showed that F.IX Nagoya seemed to be cleaved by neither F.XIa nor RVV-X, F.IX Kashihara was cleaved partially by F.XIa but not by RVV-X, and that F.IX Niigata was cleaved completely at the rate similar to normal F.IX, though the resultant product of F.IX Niigata did not show any F.IXa activity. These results favored the view that hemophilia B+ or BM is a heterogeneous disorder.
从一名柏原血友病B患者和两名BM型血友病患者的血浆中分离出异常的凝血因子IX。通过依次使用抗凝血因子IX单克隆抗体-琼脂糖凝胶、肝素-琼脂糖凝胶和二乙氨基乙基-琼脂糖凝胶A-50亲和层析，将凝血因子IX纯化至均一状态。分离出的蛋白质在分子量和交叉免疫电泳中的迁移率与正常凝血因子IX相同。通过激活因子XIa/钙离子或蛇毒凝血酶（RVV-X）/钙离子诱导纯化蛋白质的有限蛋白水解。一项时间进程研究表明，名古屋凝血因子IX似乎既不被因子XIa也不被RVV-X切割，柏原凝血因子IX被因子XIa部分切割但不被RVV-X切割，新泻凝血因子IX以与正常凝血因子IX相似的速率被完全切割，尽管新泻凝血因子IX的最终产物未显示任何凝血因子IXa活性。这些结果支持了B+型或BM型血友病是一种异质性疾病的观点。
