Rangel H A, Serafim M B, Sakurada J K
Ann Immunol (Paris). 1978 Feb-Mar;129(2-3):401-12.
The possibility of use a stable diazonium compound, to replace bisdiazotized benzidin (BDB) for conjugating antigens to red blood cells was investigated. Five different batches of 4,4'-biphenylbisdiazonium fluoroborate (BDF) were prepared and tested in conjugation experiments. The sensitivity to specific lysis of red cells coated with different antigens (EG cells) was assayed by both direct and indirect passive haemolysis tests. Optimal conditions for the preparation of EG cells with different protein antigens were established. With the exception of the concentration of the bifunctional reagent, these conditions were similar to those previously reported for BDB. The antibody content of different anti-protein sera could be determined, with a 10 % error, by using EG cells prepared under optimal conditions in indirect passive haemolysis tests. The amount of antibody detected by this method varied from 7 to 12 ng N Ab/ml, depending upon the nature of the antigen fixed to red cell. The content of BSA in dilute solutions could be estimated by the specific inhibition of the indirect passive haemolysis tests. Testing different samples of BDF maintained at different conditions showed that the compound could be kept at room temperature, during the least four years, without appreciable loss of the conjugating properties, provided it was kept in the dry state and in the dark.
研究了使用稳定的重氮化合物替代双偶氮联苯胺（BDB）将抗原与红细胞结合的可能性。制备了五批不同的4,4'-联苯双氟硼酸重氮盐（BDF）并在结合实验中进行测试。通过直接和间接被动溶血试验测定了用不同抗原包被的红细胞（EG细胞）对特异性裂解的敏感性。确定了用不同蛋白质抗原制备EG细胞的最佳条件。除双功能试剂的浓度外，这些条件与先前报道的BDB条件相似。通过在间接被动溶血试验中使用在最佳条件下制备的EG细胞，可以以10%的误差测定不同抗蛋白质血清的抗体含量。通过该方法检测到的抗体量为7至12 ng N Ab/ml，这取决于固定在红细胞上的抗原的性质。可以通过间接被动溶血试验的特异性抑制来估计稀溶液中牛血清白蛋白（BSA）的含量。对在不同条件下保存的不同BDF样品进行测试表明，只要保持干燥并避光，该化合物可以在室温下保存至少四年，而其结合特性不会有明显损失。
