Tissue plasminogen activator in human blood. 2 Gel filtration analysis of t-PA in human plasma by the method of ELISA and parabolic rate assay.:(2) Gel filtration analysis of t-PA in human plasma by the method of ELISA and parabolic rate assay
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It was reported that immediate acidification is necessary for measurement of t-PA activity in blood samples to avoid its inactivation by t-PA inhibitor (PAI).In this study, the immunological activity and enzymatic activity of t-PA were determined by ELISA method and parabolic rate assay method, respectively, and the interaction between t-PA and PAI was analysed by gel filtration method with Sephadex G-200. The following results were obtained.1. Standard t-PA, obtained from Bio pool AB, showed one peak with Mr=60kDa by gel filtration, in which both immunological and enzymatic activities were found consistently.2. By the same method, the plasma sample from a pregnant woman (PAI rich plasma) showed one peak with Mr=120kDa in which only immunological activity of t-PA was found, and no enzymatic activity was found.3. On the other hand, the mixture of standard t-PA and the plasma showed an increased peak with Mr=120kDa showing only immunological activity of t-PA, and showed a decreased peak with Mr=60kDa showing enzymatic activity of t-PA.From these results, it was presumed that activity of t-PA (Mr=60kDa) was immediately inhibited with PAI in human blood, by forming a t-PA -PAI complex (Mr=120kDa), which was undissociable under the gel filtration method. Accordingly, the Mr of PAI seemed to be 60kDa. PAI might play an important role in regulation of fibrinolysis, and its clinical significance remained to be elucidated.
- 一般社団法人 日本血栓止血学会の論文
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