Singh V
Indian J Exp Biol. 1989 Jan;27(1):1-4.
Monoclonal anti-GnRH antibodies (MoAbs) P862, P778, P813 and P764 reacted optimally to native GnRH and poorly to GnRH(OH) of sequence 4-6, 7-10, 4-10 and 1-10. The heptapeptide 4-10 showed maximum reactivity amongst the four peptides tested for immunoreactivity. Sepharose 4B-GnRH(OH)4-10 (H-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-OH) column was therefore used to purify the fraction of MoAb reacting to this sequence. The affinity purified MoAbs (A-MoAbs) were further characterized for their binding to the different sequences and affinity with native GnRH. The binding, cross-reactivity and affinity characteristics of A-MoAbs were comparable with those of MoAbs. Immunoreactivity of A-MoAbs was also observed to be partly regained when GnRH(OH)1-10 was coupled to Lys, Lys-MDP or H-Ala-Ala-Thr-Lys-Pro-Arg-OH. These observations clearly demonstrate that MoAbs were neither contaminated nor were sequence specific but were directed against the conformation of the molecule.
单克隆抗促性腺激素释放激素抗体（MoAbs）P862、P778、P813和P764对天然促性腺激素释放激素（GnRH）反应最佳，而对序列4 - 6、7 - 10、4 - 10和1 - 10的GnRH（OH）反应较差。七肽4 - 10在测试的四种免疫反应性肽中显示出最大反应性。因此，使用琼脂糖4B - GnRH（OH）4 - 10（H - Ser - Tyr - Gly - Leu - Arg - Pro - Gly - OH）柱来纯化与该序列反应的MoAb组分。对亲和纯化的单克隆抗体（A - MoAbs）与不同序列的结合及其与天然GnRH的亲和力进行了进一步表征。A - MoAbs的结合、交叉反应性和亲和力特征与MoAbs相当。当GnRH（OH）1 - 10与赖氨酸、赖氨酸 - MDP或H - Ala - Ala - Thr - Lys - Pro - Arg - OH偶联时，还观察到A - MoAbs的免疫反应性部分恢复。这些观察结果清楚地表明，MoAbs既未被污染也不是序列特异性的，而是针对分子的构象。
