Keilacker H, Dietz H, Witt S, Woltanski K P, Berling R, Ziegler M
Biomed Biochim Acta. 1986;45(8):1093-102.
We investigated four insulin-specific hybridoma antibodies with respect to their kinetic properties as well as the binding behaviour of some combinations of them. From equilibrium binding data all but one antibodies were shown to bear homogeneous binding sites. They revealed homogeneity of binding sites also by kinetic experiments, thus with high probability being monoclonal. At 0 degree C, two of them showed discrepancies between kinetic and steady state binding data in as much as, at steady state, the measured bound-to-free ratio of tracer insulin was 3-4 times lower than calculated from kinetic data. Thus a simple bimolecular reaction mechanism could possibly not be applicable. Mixing two monoclonal insulin antibodies, neither cooperative nor additive binding to the insulin molecule could be observed but only competitive effects. Especially, no positive cooperativity between two or more antibodies could be detected, which would be able to account for the higher affinity usually observed for polyclonal vs. monoclonal antibodies.
我们研究了四种胰岛素特异性杂交瘤抗体的动力学特性以及其中一些组合的结合行为。从平衡结合数据来看，除一种抗体外，其他所有抗体均显示具有均一的结合位点。通过动力学实验，它们也显示出结合位点的均一性，因此很可能是单克隆抗体。在0℃时，其中两种抗体在动力学和稳态结合数据之间存在差异，因为在稳态下，测得的示踪胰岛素的结合与游离比率比根据动力学数据计算的值低3至4倍。因此，简单的双分子反应机制可能不适用。将两种单克隆胰岛素抗体混合后，未观察到对胰岛素分子的协同或加性结合，仅观察到竞争效应。特别是，未检测到两种或更多种抗体之间的正协同作用，而这种正协同作用本可解释通常观察到的多克隆抗体与单克隆抗体相比具有更高亲和力的现象。
