PKA活性测定——微量滴定板方法的探讨

Determination of PKA Activity by Microtiter Plate Method

本文应用微量滴定板方法测定血液制品中残留的PKA活性.结果表明,用0.05M Tris/0.15M NaCl,pH8.0缓冲液,PKA和PK在室温下培育30分钟,然后加入激肽释放酶显色底物,在室温下再继续培育15分钟是适宜的.该方法重复性好,所测得IVIG中PKA的含量与其生产工艺、大白鼠降压实验相符,并与用芬兰红十字血液中心方法测得的结果基本一致.

In the paper, a microtiter plate method was used to determine the residual prekallikrein activator(PKA) activity in blood products.The result of study indicated that it would be satisfactory that the reaction mixture of PKA and prekallikrein (PK) was iucul ?? at room temperature for 30 minutes with 0.05M Tris/0.15M NaCl buffer(pH 8.0) , and then the mixture continued to be incubated at room temperature for another 15 minutes after the chromogenic substrate for kallikrein was added. The method had a good reproducibility. The PKA contents determined in IVIG agreed with that of the processing method of IVIG and the hypotensive test of rats, and were basically consistent with the result, determined by the method developed by the Finnish Red Cross Blood Transfusion Service.