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以三硝基苯磺酸钠(TNBS)修饰铁传递蛋白(Tf)及BSA,并制得高效价的特异性抗TNP血清。运用放射受体分析法比较Tf经TNP修饰前后与人肝癌培养细胞膜上Tf受体(TR)的亲和力,证明Tf经TNP化修饰后未改变其与受体的亲和力。应用TNP-Tf、抗TNP血清为第一抗体的间接酶标法检测人肝癌培养细胞膜上的TR,其检测敏感度是应用Tf及抗TNP血清为第一抗体的间接酶标法的4倍。以硝酸-醋酸纤维素膜作载体,用间接酶标法进行定量比较Tf及TNP-Tf的最小检测量,则Tf的最小检测限为5ng,而TNP-Tf的最小检测限为1ng。
The transferrin (Tf) and BSA were modified by sodium trinitrobenzenesulfonate (TNBS), and high titer anti-TNP serum was obtained. The binding affinity of Tf to Tf receptor (TR) on cultured human hepatocellular carcinoma cell membrane before and after TNP modification was compared by radioimmunoassay. The results showed that Tf did not change its affinity with receptor after modified by TNP. TNP-Tf and TNP sera were used as the primary antibody to detect the TR in human hepatocellular carcinoma cell membrane. The detection sensitivity was four times higher than that of indirect ELISA using Tf and anti-TNP serum as the primary antibody. Using nitric acid-cellulose acetate membrane as carrier, the minimum detection limit of Tf and TNP-Tf were compared quantitatively by indirect enzyme-labeling method. The minimum detection limit of Tf was 5ng, while the minimum detection limit of TNP-Tf was 1ng.