[prekallikrein浓缩物的制备及其在血液衍生物中激活剂的测定]。

Ceskoslovenska farmacie Pub Date : 1992-02-01
E Simonianová
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引用次数: 0

摘要

测定从血浆制备的制剂中哈格曼因子(HFf)片段的增加水平可作为其临床应用中潜在反应性的指标。证明HFf的存在是基于这样一个事实,即含有该因子的制剂能够激活预钾likrein底物。当prekallikrein浓度足够时,形成的活性酶kallikrein的量与HFf的量成正比。通过切割特定的显色底物no - pro - ph - arg - pna来确定钾激肽的量。盐的存在对反应激活阶段的速率有负面影响。所描述的从人血浆中制备prekallikrein浓缩物的方法是对Levy和Sober分离人免疫球蛋白部分的方法的改进。加入六己甲醚的血浆在表面完整的容器中用0.0175 mol/l pH为6.3的磷酸缓冲液与六己甲醚溴化物平衡的DEAE Sephadex上吸附。在上述条件下,Prekallikrein和IgG不吸附在阴离子交换器上。所得上清液可用作预钾化酶底物,测定血液衍生物中HFf活性。
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[Preparation of a prekallikrein concentrate and determination of its activator in blood derivatives].

The determination of an increased level of a fragment of the Hageman's factor (HFf) in preparations prepared from blood plasma can be an index of their potential reactivity in clinical application. The demonstration of the presence of HFf is based on the fact that the preparation containing this factor is able to activate the prekallikrein substrate. When the concentration of prekallikrein is sufficient, the amount of the developed active enzyme kallikrein is proportional to the amount of HFf. The amount of kallikrein is determined by cleaving the specific chromogenic substrate NO-Pro-Phe-Arg-pNA. The presence of salts negatively influences the rate of the activating stage of the reaction. The described preparation of the prekallikrein concentrate from the human blood plasma is a modification of the method of Levy and Sober) for the separation of the human immunoglobulin fraction. The blood plasma with an addition of hexadimethrinebromide is sorbed on DEAE Sephadex equilibrated with 0.0175 mol/l phosphate buffer pH 6.3 with hexadimethrinebromide in a vessel with an intact surface. Prekallikrein together with IgG are not sorbed on the anion exchanger under the above-mentioned conditions. The obtained supernatant can be employed as the prekallikrein substrate to determine the HFf activity in blood derivatives.

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