Markarian A N, Cherednikova T V, Dzantiev B B
Bioorg Khim. 1986 Mar;12(3):327-31.
A modified procedure has been worked out for preparing a conjugate of porcine insulin with E. coli beta-galactosidase employing a heterobifunctional reagent, N-hydroxysuccinimidyl m-maleimidobenzoate. Optimal conditions for insulin acylation and subsequent coupling with beta-galactosidase were selected that afforded the conjugate in a high yield. The ability of the modified antigen to react with antibody was evaluated in the reaction of conjugate binding with immobilized monoclonal antibody to insulin. The conjugate almost completely retained the enzymatic activity and reacted with high specificity with the antibody to insulin. The conjugate can be used in competitive ELISA of insulin.
已制定出一种改良方法，使用异双功能试剂间马来酰亚胺苯甲酸N-羟基琥珀酰亚胺酯制备猪胰岛素与大肠杆菌β-半乳糖苷酶的偶联物。选择了胰岛素酰化及随后与β-半乳糖苷酶偶联的最佳条件，从而以高产率获得偶联物。在偶联物与固定化抗胰岛素单克隆抗体的结合反应中，评估了改良抗原与抗体反应的能力。该偶联物几乎完全保留了酶活性，并且与抗胰岛素抗体具有高特异性反应。该偶联物可用于胰岛素的竞争性酶联免疫吸附测定。
