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本工作合成了两个标记抗原:孕酮-6-巯基乙酸-~(125)I-酪氨(P-6-CMTE-[~(125)I]-T_(yγ))和孕酮-16-巯基乙酸-~(125)I-一组氨(P-16-CMTE-[~(125)I]-His)。两标记抗原分别与免疫原孕酮-6-疏基乙酸-BSA(P-6-CMTE-BSA)的抗体和免疫原孕酮-112-半琥珀酸-BSA(P-112-HS-BSA)的抗体做同桥、同位;同桥、异位;异桥、异位的结合实验。还对C-16位标记物做了两相标记条件的研究。实验结果表明:(1)巯基乙酸桥是一个弱极性的、三原子组成的直链桥,具有弱的免疫原性。联接巯基乙酸桥,对孕酮半抗原分子性质影响不大。C-6位标记物与同桥、同位的C-6位抗体结合的结果类似于未接任何配体的~3H-孕酮标记抗原,与异桥、异位的C-11位抗体有结合。(2)标记物和免疫原与配体的偶联部位,要避开甾体分子的功能部位,以致不影响甾体的结构特征,使抗体识别能力增强。D环是孕酮分子的功能部位,在D环
In this work, we synthesized two labeled antigens: progesterone-6-thioglycolic acid- 125-I tyrosine (P-6-CMTE- [~ (125) I] -Tγ (yγ) - thioglycolic acid - ~ (125) I - a group of ammonia (P-16-CMTE- [~ (125) I] -His). Two labeled antigens were immunoprecipitated with the progesterone-6-spikylacetic acid-BSA (P-6-CMTE-BSA) Of the antibodies do the same bridge, the same bit; with the bridge, ectopic; different bridge, ectopic binding experiments. A two-phase labeling condition was also studied for the C-16 marker. The experimental results show that: (1) thioglycolic acid bridge is a weakly polar, three-atom linear bridge composed of a weak immunogenicity. Linking thioglycolic acid bridge has little effect on the molecular properties of progesterone hapten. The binding of the C-6 marker to the isostere, C-6 antibody at the same position was similar to that of the ~ 3H-progesterone-labeled antigen without any ligand and was found to bind to the isoproterenol and C-11 antibody . (2) The conjugate between the label and the immunogen and the ligand should avoid the functional site of the steroid molecule, so as not to affect the structural characteristics of the steroid and enhance the recognition ability of the antibody. Ring D is the functional site of progesterone molecules in the D ring