Isolation of Human Fibrinogen and its Derivatives by Affinity Chromatography on Gly-Pro-Arg-Pro-Lys-Fractogel
Isolation of Human Fibrinogen and its Derivatives by Affinity Chromatography on Gly-Pro-Arg-Pro-Lys-Fractogel
复制标题
通过 Gly-Pro-Arg-Pro-Lys-Fractogel 亲和色谱分离人纤维蛋白原及其衍生物
作者：
C. Kuyas;A. Haeberli;P. Walder;P. Straub
通过固定化合成纤维蛋白α-链n端GlyProArg的五肽GlyProArgProLys，建立了一种新的从抗凝血浆中定量分离纤维蛋白原的亲和方法。事实证明，该方法在易用性和产量方面优于所有已知的分离方法，因为与免疫可测量的纤维蛋白原量相比，纤维蛋白原可以一步从血浆中分离出来，回收率超过95%。此外，纤维连接蛋白、因子XIII或纤溶酶原等污染蛋白的数量可以忽略不计，聚丙烯酰胺凝胶电泳检测分离的纤维蛋白原纯度高于95%。吸附性达90%以上。这种亲和纯化方法的另一个优点是可以从小血浆样品（< 5ml）中定量分离纤维蛋白原。此外，异常纤维蛋白原分子，只要其GlyProArg的互补结合位点被保留，也可以定量分离，与正常纤维蛋白原相比，没有任何溶解度差异。此外，源自质消化的纤维蛋白（ogcn）片段可以根据其对GlyProArg的亲和力进行分离。所述亲和凝胶可使用50次以上而不损失容量。
Summary With an immobilized synthetic pentapeptide GlyProArgProLys comprising the N-terminal sequence GlyProArg of the α-chain of fibrin, a new affinity method for the quantitative isolation of fibrinogen out of anticoagulated plasma was developed. The method proved to be superior to all known isolation methods in respect to ease of use and yield, since fibrinogen could be isolated in one step out of plasma with a recovery of more than 95% when compared to the immunologically measurable amounts of fibrinogen. Moreover the amounts of contaminating proteins such as fibronectin, factor XIII or plasminogen were negligible and the purity of the isolated fibrinogen was higher than 95% as measured by polyacrylamide gel electrophoresis. The clottability was 90% and more. Another advantage of this affinity purification method is the possibility to isolate fibrinogen quantitatively out of small plasma samples (<5 ml). Further, abnormal fibrinogen molecules, provided their complementary binding site for GlyProArg is preserved, may also be quantitatively isolated independent of any solubility differences as compared to normal fibrinogen. In addition fibrin(ogcn) fragments originating from plasmic digestion can be separated on the basis of their affinity to GlyProArg. The described affinity gel can be used more than 50 times without any loss of capacity.
人纤维蛋白原伽玛链中聚合和钙结合所必需的片段的定位。
影响因子：
2.9
作者：
Váradi,A;Scheraga,HA
通讯作者：
Scheraga,HA