【摘要】 为了提高放射免疫分析法（RIA）的灵敏度,达到度量人外周血浆中血管紧张素Ⅱ（ATII）的要求,必需采用亲和力特别高的特异性抗ATII 抗血清。我们使用不同的载体和结合方法,制备了五种ATIIa 免疫原,以提高其抗原性。它们是:1)聚乙烯吡咯酮吸附血管紧张素II 酰胺（ATIIa）;2)碳二亚胺缩合兔血清白蛋白与ATIIa;3)戊二醛（GDA）结合牛血清白蛋白（BSA）与ATIIa;4)GDA 结合多聚赖氨酸（PL）与ATIIa;5)碳二亚胺缩合琥珀酰化多聚赖氨酸（PL-Succ）与ATIIa。在分组免疫家兔（皮内多点或多途径注射）后,用生物鉴定、被动血球凝集试验和放射免疫稀释曲线法检测及跟踪抗血清的抗ATII 效价。并用RIA 法以Scatchard 图求亲和常数K_?统计显示在组间存在显著差别,以BSA-GDA-ATIIa 及PL-Succ-ATIIa 效果为佳。在3～5个月内成功地获得了K_a>10¹⁰升/克分子,适用于ATII 的放射免疫测定的一些抗血清。其中之一已经满意地用于建立一个特异、灵敏的RIA,可以直接测定人体外周血中低浓度的ATII。另一些也有相似的能力。对工作中涉及一些问题,如免疫原制备、影响抗血清质量的因素以及抗ATII 检测方法等,作了初步的比较和讨论。本试验结果,从抗原消耗量、抗体产生速度、成功率及抗血清的效价和亲和力等方面,看来较某些报告有优越之处。更多还原

【Abstract】 There is much need for specific and high-affinity antisera to angiotensin Ⅱ（ATⅡ）for the radioimmunoassay of AT Ⅱ concentration in human peripheral blood.Fivecoupled immunogens were prepared by using different carriers and methods of couplingto render AT Ⅱ amide（AT Ⅱa）antigenic.They are:1. ATIIa adsorption to polyvinylpyrrolidone.2.Condensation of ATIIa with rabbit serum albumin by carbodiimide reagent.3.Conjugation of AT Ⅱa wish bovine serum albumin（BSA）via glutaricdialdehyde（GDA）.4.Conjugation of AT Ⅱa with poly-L-lysine via GDA.5.Condensation of AT Ⅱa with succinylated poly-L-lysine（PL-Succ）bycarbodiimide reagent.Groups of rabbits were immunized by multiple-site intradermal injection orthrough multiple routes.The potency of anti-AT Ⅱ of the various antisera weredetected and followed by biological assay,passive hemagglutination test and RIA.Theaffinity constants（K_a）of these antisera to AT Ⅱa were also determined by Scatchardplotting using RIA.Among groups,different effects were shown by statistical analysis.The BSA-GDA-ATIIa and the PL-Succ-AT Ⅱa were shown to be highly effective.Certain antisera,with K_a values of 10¹⁰l/mol and suitable for use in RIA,have been obtained 3～5months after immunization.One of these has been successfully employed in a specificand sensitive RIA for the direct measurement of small concentrations of ATIIa inhuman peripheral blood.In connection with our experiments relating to the preparations of immunogens,the methods of detecting antibody and the factors affecting the qualities of antiserawere compared and discussed.Our results,as compared with most other publishedreports,offer some distinct advantages.更多还原