Delcros J G, Clement S, Thomas V, Quemener V, Moulinoux J P
Laboratoire d'Immunochimie INSERM, Faculté de Médecine Lyon Sud, Oullins, France.
J Immunol Methods. 1995 Sep 25;185(2):191-8. doi: 10.1016/0022-1759(95)00114-p.
The reactivity of an anti-spermine MAb (SPM8-2) toward polyamines either free or bound to a solid surface was investigated using equilibrium dialysis and ELISA methods. When polyamines were covalently linked to hydrophilized microtiter plates using carbodiimide, the MAb SPM8-2 reacted both with spermine and spermidine, with a higher affinity for the latter, but did not show any reactivity towards bound putrescine. In contrast, the MAb SPM8-2 reacted with all three polyamines bound to the microtiter plates with glutaraldehyde, with an affinity in the order: putrescine > spermidine > spermine. Equilibrium dialysis and competitive ELISA tests showed that the MAb SPM8-2 exhibited high affinity for free spermine and 50% and 5% cross-reactivity with free spermidine and putrescine respectively. The affinity of the MAb SPM8-2 for putrescine, spermidine and spermine appears to depend on whether the polyamine is free or bound. The antigenicity of the polyamines differs according to the nature of their link to the solid phase. These observations are discussed in the light of the structural modification produced by covalent binding of the polyamines. It is also concluded that when antibodies are used, due care has to be exercised in choosing the appropriate immunoassay for determining the specificity of antibodies directed against small haptens such as the polyamines.
使用平衡透析法和酶联免疫吸附测定（ELISA）法研究了抗精胺单克隆抗体（SPM8 - 2）对游离或结合于固体表面的多胺的反应性。当使用碳二亚胺将多胺共价连接到亲水化的微量滴定板上时，单克隆抗体SPM8 - 2与精胺和亚精胺均发生反应，对后者的亲和力更高，但对结合的腐胺未表现出任何反应性。相比之下，单克隆抗体SPM8 - 2与通过戊二醛结合到微量滴定板上的所有三种多胺都发生反应，亲和力顺序为：腐胺＞亚精胺＞精胺。平衡透析和竞争性ELISA试验表明，单克隆抗体SPM8 - 2对游离精胺具有高亲和力，对游离亚精胺和腐胺的交叉反应性分别为50%和5%。单克隆抗体SPM8 - 2对腐胺、亚精胺和精胺的亲和力似乎取决于多胺是游离的还是结合的。多胺的抗原性因其与固相连接的性质而异。根据多胺共价结合产生的结构修饰对这些观察结果进行了讨论。还得出结论，当使用抗体时，在选择合适的免疫测定方法以确定针对多胺等小分子半抗原的抗体特异性时必须格外小心。
