Shankar H, Senatore F, Zuniga P, Venkataramani E
J Biomed Mater Res. 1987 Jul;21(7):897-912. doi: 10.1002/jbm.820210706.
Plasmin was immobilized on collagenous substrates using carbodiimide as a linking agent. The kinetics of soluble and immobilized plasmin were monitored by reacting them with the chromogenic substrate S-2251 (H-D-Val-Leu-Lys-pNA) in the presence and absence of a2-antiplasmin (a2-PI). The ability of immobilized plasmin to lyse synthetic clots formed from fibrinogen and thrombin was determined by detecting the formation of fibrin degradation products (FDP). The activity of immobilized plasmin was 0.02 casein units (CU)/mg of collagen. The kinetic analysis of soluble and immobilized plasmin in the presence and absence of a2-PI shows that while soluble plasmin activity was inhibited by the presence of a2-PI, the plasmin inhibitor did not interfere with the ability of immobilized plasmin to attack fibrin. In the absence of a2-PI, the ability of the immobilized plasmin to lyse synthetic clots was the same as that of soluble plasmin. In the presence of a2-PI, immobilized plasmin produced twice the amount of FDP as did soluble plasmin. The immobilized plasmin activity was stable for a period of at least 3 months.
使用碳二亚胺作为连接剂将纤溶酶固定在胶原底物上。通过在存在和不存在α2 - 抗纤溶酶（α2 - PI）的情况下使可溶性和固定化纤溶酶与发色底物S - 2251（H - D - Val - Leu - Lys - pNA）反应，监测其动力学。通过检测纤维蛋白降解产物（FDP）的形成来确定固定化纤溶酶溶解由纤维蛋白原和凝血酶形成的合成凝块的能力。固定化纤溶酶的活性为0.02酪蛋白单位（CU）/毫克胶原。在存在和不存在α2 - PI的情况下对可溶性和固定化纤溶酶的动力学分析表明，虽然α2 - PI的存在抑制了可溶性纤溶酶的活性，但纤溶酶抑制剂并不干扰固定化纤溶酶攻击纤维蛋白的能力。在不存在α2 - PI的情况下，固定化纤溶酶溶解合成凝块的能力与可溶性纤溶酶相同。在存在α2 - PI的情况下，固定化纤溶酶产生的FDP量是可溶性纤溶酶的两倍。固定化纤溶酶的活性至少在3个月内保持稳定。
