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目的探讨微孔板酶联比色测定法在蚯蚓粗提物蛋白质量浓度、蚓激酶活性测定中的应用。方法蛋白质量浓度采用改良Lowry法,蚓激酶活性采用TAME法,分别使用常量体积和微量体积,用紫外-可见分光光度计和酶标检测仪进行测量,用SPSS软件进行直线拟合,做出标准曲线,比较测量结果。结果标准曲线截距RSD<5%,用标准曲线检测BSA的结果显示,2种质量浓度、2种方法的回收率分别为99.6%,94.6%,101.2%和96.5%,RSD分别为3.30%,5.05%,3.36%和2.59%,经t检验3种检测结果差异无统计学意义,可以认为2种检测方法测得的数据一致。结论可以采用微量法进行蚯蚓粗提物蛋白质量浓度、蚓激酶活性的测定。
Objective To investigate the application of microtiter plate ELISA in the determination of protein concentration and lumbrokinase activity of earthworm crude extract. Methods The protein concentration was modified by Lowry method and the activity of lumbrokinase was determined by TAME method. The volume and volume were determined by UV-visible spectrophotometer and enzyme-labeled detector, and fitted by SPSS software to make the standard Curve, compare measurement results. Results The results showed that the RSD of standard curve was less than 5%. The results of BSA with standard curve showed that the recovery rates of two kinds of mass concentration and two methods were 99.6%, 94.6%, 101.2% and 96.5%, RSD were 3.30% 5.05%, 3.36% and 2.59%, respectively. There was no significant difference between the three test results by t test, which means that the data obtained by the two test methods are consistent. Conclusion Micropore method can be used to determine the protein concentration of earthworm crude extract and the activity of lumbrokinase.