Studies of hypersensitivity to low molecular weight substances. III. The 2,4-dinitrophenyl group as a determinant in the preciptin reaction.
Studies of hypersensitivity to low molecular weight substances. III. The 2,4-dinitrophenyl group as a determinant in the preciptin reaction.
复制标题
对低分子量物质过敏的研究。
作者：
H. Eisen;M. E. Carsten;S. Belman
摘要2，4-二硝基苯基通过偶氮键或通过取代游离氨基与蛋白质结合，已被研究为沉淀素反应中的决定簇。所获得的沉淀素曲线类似于高分子量纯化抗原所遇到的沉淀素曲线，主要例外是在抗体区域中过量抗原没有完全沉淀。二硝基苯基偶氮卵白蛋白是一个不太有效的沉淀剂比共轭卵白蛋白，其中二硝基苯基取代的游离氨基。然而，前者是相对更有效地沉淀抗体从抗血清二硝基苯偶氮蛋白比从抗血清制备对氨基取代的二硝基苯蛋白。二硝基苯-牛γ球蛋白混合抗血清的球蛋白组分至少含有三种抗体，即牛γ球蛋白沉淀的抗体、二硝基苯-卵清蛋白沉淀的抗体和仅二硝基苯-牛γ球蛋白沉淀的抗体。这些分别以1：1：1.3的比例存在。在用这些抗原中的每一种进行最大沉淀后，通过平衡透析法测定e-N-二硝基苯赖氨酸的上清液9结合，e-N-二硝基苯赖氨酸是一种半抗原，其中二硝基苯与赖氨酸以与免疫缀合物相同的方式结合。三种情况下的结合相对于未吸收的免疫球蛋白的结合的比例分别为1：0.6：0.25。因此，牛γ球蛋白沉淀的抗体对二硝基苯基没有可检测的特异性，而异源偶联物（二硝基苯基-卵清蛋白）沉淀的抗体对二硝基苯基决定簇的亲和力大于沉淀完全同源偶联物（二硝基苯基-牛γ球蛋白）所需的抗半抗原抗体。具有双重特异性的抗体，对半抗原具有一种特异性，而对免疫缀合物的蛋白质具有明显不同的特异性。
Summary The 2,4-dinitrophenyl group, combined with proteins through azo linkage or by substitution in free amino groups, has been studied as a determinant in the precipitin reaction. The precipitin curves obtained resemble those encountered with purified antigens of high molecular weight with the chief exception that in the region of antibody excess antigen is not completely precipitated. Dinitrophenyl-azo-ovalbumin was a less effective precipitant than the conjugated ovalbumin in which dinitrophenyl groups were substituted in free amino groups. The former, however, was relatively more effective in precipitating antibody from antisera to dinitrophenyl-azo-proteins than from antisera prepared against amino-substituted dinitrophenyl-proteins. The globulin fraction of pooled antisera to dinitrophenyl-bovine γ globulin contained at least three species of antibodies—viz: those precipitated by bovine γ globulin; those precipitated by dinitrophenyl-ovalbumin and those precipitated only by dinitrophenyl-bovine γ globulin. These were present in the ratio of 1:1:1.3, respectively. After maximal precipitation with each of these antigens the supernates9 binding of e-N-dinitrophenyllysine, a hapten in which dinitrophenyl is combined with lysine in the same way as in the immunizing conjugate, was determined by the method of equilibrium dialysis. The binding in the three cases, relative to the binding by the unabsorbed immune globulin, was in the ratio of 1:0.6:0.25, respectively. Hence, the antibodies precipitated by bovine γ globulin had no detectable specificity for the dinitrophenyl group, and the antibodies precipitated by the heterologous conjugate (dinitrophenyl-ovalbumin) had a greater affinity for the dinitrophenyl determinant than those antihapten antibodies that required for precipitation the fully homolgous conjugate (dinitrophenyl-bovine γ globulin). Antibodies with dual specificity, i.e., with one specificity for the hapten and a distinctly different one for the protein of the immunizing conjugate, were not detected.