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采用比较简便、有效的提取方法,从150g人心肌中得到660mg肌球蛋白粗品,经DEAE-Sephadex A-50纯化后得到纯肌球蛋白320mg,比活力达5.05u/mg蛋白,酶活力回收36％。进一步以纯肌球蛋白制得电泳纯的心肌肌球蛋白轻链,并以之免疫新西兰兔而获得抗人心肌肌球蛋白轻链的抗血清。抗血清具有较好的特异性,亲和常数为1.16×10~9M~(-1);当稀释度为1:2000时,可用于酶联免疫吸附分析。此外,还测定了心肌肌球蛋白及其轻链成份的理化特性和酶反应动力学特征。
Using simple and effective extraction method, 660mg of myosin crude was obtained from 150g of human myocardium and pure myosin 320mg was obtained after purification by DEAE-Sephadex A-50. The specific activity reached 5.05u / mg protein, and the enzyme activity recovery 36 %. Further, pure electrophoresis of myocardium myosin light chain was made by pure myoglobin, and antisera of anti-human myocardial myosin light chain was obtained by immunizing New Zealand rabbit. The antiserum has good specificity and the affinity constant is 1.16 × 10 ~ 9M ~ (-1). When the dilution is 1: 2000, it can be used for enzyme-linked immunosorbent assay. In addition, the physicochemical properties and enzymatic kinetic characteristics of myocardial myosin and its light chain components were also determined.