Singh V
Biochem Cell Biol. 1986 Dec;64(12):1372-7. doi: 10.1139/o86-180.
Immunochemical studies on monoclonal and conventional anti-LHRH antibodies have been reported. The association constants (Ka) were in the order of 10(9)-10(10) L/mol when calculated from Scatchard and Steward-Petty plots. Heterogeneity indices (a) calculated from Sips plot were nearly 1.0, indicating the homogeneous nature of monoclonals. Most of the conventional anti-LHRH produced by monkey, baboon, rabbit, and dog, by immunization using LHRH linked to tetanus toxoid by the carbodiimide condensation method, showed a single slope in Scatchard analysis (except two dog antisera) and a values were nearly 1.0. Monoclonals and conventionals reacted most strongly with native LHRH(NH2). Monoclonals showed poor reactivity with LHRH free acid and LHRH fragments containing free acid. The C-terminus tetrapeptide 7-10 showed 10 times more reactivity than tripeptide 4-6. The heptapeptide 4-10 showed 100 and 1000 times more reactivity than the tetra- and tri-peptide, respectively. Introduction of the tripeptide pGlu-His-Trp-OH to heptapeptide 4-10 caused five times more inhibition in reactivity than the heptapeptide. Conventional anti-LHRH antibodies manifested specificity to the C-terminus of LHRH. These sera did not react with LHRH free acid and LHRH fragments of sequence 4-6, 7-10, and 4-10. The complete loss of reactivity of conventional antibodies and poor reactivity of monoclonal antibodies was partly regained when LHRH free acid was coupled to Lys, Lys-MDP, or (Ala)2-tuftsin, suggesting that for monoclonals and conventionals the antigenic determinant was confined to the conformation involving the C-terminus amide of LHRH.
关于单克隆和传统抗促黄体激素释放激素（LHRH）抗体的免疫化学研究已有报道。根据Scatchard和Steward - Petty图计算，其结合常数（Ka）约为10⁹ - 10¹⁰L/mol。从Sips图计算出的异质性指数（a）接近1.0，表明单克隆抗体具有均一性。通过碳二亚胺缩合法将LHRH与破伤风类毒素连接后免疫猴、狒狒、兔和狗产生的大多数传统抗LHRH抗体，在Scatchard分析中显示单一斜率（除两份狗抗血清外），a值接近1.0。单克隆抗体和传统抗体与天然LHRH（NH₂）反应最强。单克隆抗体与游离酸形式的LHRH及含游离酸的LHRH片段反应性较差。C末端四肽7 - 10的反应性比三肽4 - 6高10倍。七肽4 - 10的反应性分别比四肽和三肽高100倍和1000倍。将三肽pGlu - His - Trp - OH引入七肽4 - 10会使反应性抑制比七肽高5倍。传统抗LHRH抗体对LHRH的C末端表现出特异性。这些血清不与游离酸形式的LHRH及序列4 - 6、7 - 10和4 - 10的LHRH片段反应。当游离酸形式的LHRH与赖氨酸、赖氨酸 - MDP或（丙氨酸）₂ - 促吞噬肽偶联时，传统抗体完全丧失的反应性和单克隆抗体较差的反应性部分得以恢复，这表明对于单克隆抗体和传统抗体而言，抗原决定簇局限于涉及LHRH C末端酰胺的构象。
