将因子IX的首个表皮生长因子样结构域替换为因子VII的该结构域可增强体外活性及在犬血友病B模型中的活性。
Replacing the first epidermal growth factor-like domain of factor IX with that of factor VII enhances activity in vitro and in canine hemophilia B.
作者信息
Chang J Y, Monroe D M, Stafford D W, Brinkhous K M, Roberts H R
机构信息
Center for Thrombosis and Hemostasis, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-7035, USA.
出版信息
J Clin Invest. 1997 Aug 15;100(4):886-92. doi: 10.1172/JCI119604.
Using the techniques of molecular biology, we made a chimeric Factor IX by replacing the first epidermal growth factor-like domain with that of Factor VII. The resulting recombinant chimeric molecule, Factor IXVIIEGF1, had at least a twofold increase in functional activity in the one-stage clotting assay when compared to recombinant wild-type Factor IX. The increased activity was not due to contamination with activated Factor IX, nor was it due to an increased rate of activation by Factor VIIa-tissue factor or by Factor XIa. Rather, the increased activity was due to a higher affinity of Factor IXVIIEGF1 for Factor VIIIa with a Kd for Factor VIIIa about one order of magnitude lower than that of recombinant wild-type Factor IXa. In addition, results from animal studies show that this chimeric Factor IX, when infused into a dog with hemophilia B, exhibits a greater than threefold increase in clotting activity, and has a biological half-life equivalent to recombinant wild-type Factor IX.
利用分子生物学技术，我们通过将第一个表皮生长因子样结构域替换为因子VII的结构域，制备了一种嵌合因子IX。所得的重组嵌合分子，即因子IXVIIEGF1，在一期凝血试验中的功能活性与重组野生型因子IX相比至少增加了两倍。活性增加并非由于被活化的因子IX污染，也不是由于因子VIIa-组织因子或因子XIa激活速率增加所致。相反，活性增加是由于因子IXVIIEGF1对因子VIIIa具有更高的亲和力，其对因子VIIIa的解离常数（Kd）比重组野生型因子IXa低约一个数量级。此外，动物研究结果表明，这种嵌合因子IX注入患血友病B的狗体内时，凝血活性增加超过三倍，并且其生物半衰期与重组野生型因子IX相当。
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