Chamat S, Hoebeke J, Strosberg A D
J Immunol. 1984 Sep;133(3):1547-52.
After somatic cell fusion between splenocytes of immunized BALB/c mice and NS-1 myeloma cells, eight clones were obtained secreting anti-alprenolol antibodies as characterized by means of an ELISA. Four of these were subcloned and were studied further. The association constant for alprenolol ranged from 1.9 X 10(6) M-1 to 24 to 10(6) M-1. Competitive inhibition of [3H]-l-dihydroalprenolol binding revealed cross-reactivity with beta-adrenergic ligands, with a higher avidity for antagonists than for agonists. Two of the antibodies had a higher affinity for the l-isomer than for the d-isomer. The most stereospecific of these antibodies showed only affinity for beta-adrenergic antagonists and for the agonist isoproterenol. The other recognized both beta-adrenergic antagonists and agonists; it also showed an increase in tryptophan fluorescence after ligand binding. This property was used for the physicochemical study of the hapten-antibody interaction.
将免疫的BALB/c小鼠脾细胞与NS-1骨髓瘤细胞进行体细胞融合后,通过酶联免疫吸附测定法(ELISA)鉴定,获得了8个分泌抗阿普洛尔抗体的克隆。其中4个进行了亚克隆并进一步研究。阿普洛尔的结合常数范围为1.9×10⁶ M⁻¹至24×10⁶ M⁻¹。[³H]-l-二氢阿普洛尔结合的竞争性抑制显示与β-肾上腺素能配体有交叉反应,对拮抗剂的亲和力高于对激动剂的亲和力。其中两种抗体对l-异构体的亲和力高于对d-异构体的亲和力。这些抗体中立体特异性最强的仅对β-肾上腺素能拮抗剂和激动剂异丙肾上腺素具有亲和力。另一种既识别β-肾上腺素能拮抗剂又识别激动剂;它在配体结合后色氨酸荧光也增强。这一特性被用于对半抗原-抗体相互作用进行物理化学研究。