Raghava G P, Goel A, Singh A M, Varshney G C
Institute of Microbial Technology, Chandigarh, India.
Biotechniques. 1994 Dec;17(6):1148-53.
A simple microassay and computer program are described for determining the erythrocyte hemolytic potency of drugs in vitro. This microassay is sensitive for both micro as well as macro ranges of hemoglobin concentration. An ELISA reader has been adapted to read erythrocyte lysis (hemolysis), which reduces the number and culture of replicates. A computer program was developed that calculates parameters such as C50 (concentration of drug causing 50% hemolysis), C100 (concentration of drug causing 100% hemolysis) and beta (slope of the curve) and graphically expresses the hemolytic patterns of various drugs simultaneously. The program can obtain optical densities directly from a 96-well plate ELISA reader by interfacing the microplate reader to the computer or by using a keyboard. This method is useful for screening a large number of hemolytic drugs and requires lower amounts of test compounds. It may also be applicable to quantitative functional assays, such as complement-mediated hemolysis and enumeration of antibody-secreting cells. The program can be obtained from the authors on request.
本文描述了一种用于体外测定药物红细胞溶血效力的简单微量测定法及计算机程序。该微量测定法对血红蛋白浓度的微观和宏观范围均敏感。已对酶联免疫吸附测定（ELISA）读数器进行改造以读取红细胞裂解（溶血）情况，这减少了重复实验的次数和培养量。开发了一个计算机程序，可计算诸如C50（引起50%溶血的药物浓度）、C100（引起100%溶血的药物浓度）和β（曲线斜率）等参数，并同时以图形方式表达各种药物的溶血模式。该程序可通过将酶标仪与计算机连接或使用键盘直接从96孔板ELISA读数器获取光密度。此方法有助于筛选大量溶血药物，且所需测试化合物量较少。它也可能适用于定量功能测定，如补体介导的溶血和抗体分泌细胞计数。可应要求向作者获取该程序。
