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作者自蜱浸出液经Sephadex G-50凝胶过滤,离子交换层析和反向高效液相层析(RP-HPLC)分离,提炼出一种纯蛋白制品一纯粹蜱抗凝肽(TAP)。对纯化TAP抑制特性的研究表明,TAP对因子Xa活性的抑制作用与其浓度有关,抑制常数Ki=0.588±0.054nM,并且至少预孵育15分钟才能发挥最大抑制作用。因此,TAP属于慢作用、紧密结合的抑制剂。此外,TAP在体外试管内也具有抗凝活性,能延长凝血酶原时间、白陶土部份凝血活酶时间和改良Stypven时间,并与其浓度相关。
The authors extracted a pure protein product, a pure tick anti-thromboplastin (TAP), from tick tick leachate by Sephadex G-50 gel filtration, ion-exchange chromatography and reverse-phase high performance liquid chromatography (RP-HPLC). Studies on the inhibitory properties of purified TAP showed that the inhibitory effect of TAP on factor Xa activity was related to its concentration, with the inhibition constant Ki = 0.588 ± 0.054nM, and the maximum inhibitory effect could be achieved by at least 15 minutes preincubation. Therefore, TAP is a slow-acting, tight-binding inhibitor. In addition, TAP also has anticoagulant activity in vitro and prolongs and correlates with prothrombin time, kaolin partial thromboplastin time, and modified Stypven time.