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Radioimmunoassay for the Antigenic Determinants of Cholera Toxin and Its Components

机译：放射免疫法测定霍乱毒素及其成分的抗原决定簇

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A radioimmunoassay procedure is described for the detection of cholera toxin and its component polypeptide chains. Cholera toxin, A subunit, B subunit, α chain, and γ chain were iodinated by the chloramine T procedure. Radiolabeling did not significantly alter the polyacrylamide electrophoretic migration patterns of the toxin or its components. Moreover, radiolabeled toxin, B subunit, and α chain preparations retained substantial ability to bind to intestinal mucosal homogenates. The minimal amount of antitoxin detectable with radiolabeled toxin was 0.04 antitoxin units/ml. Substitution of radiolabeled B subunit, A subunit, and α chain for radiolabeled toxin decreased the sensitivity of the test. Radiolabeled γ chain did not bind to the antitoxin preparation. Competitive inhibition studies, with titrated anti-choleragen serum and radiolabeled toxin or components, indicated that the minimum concentration of toxin detectable was 7.0 × 10?8 μmol/ml at a 90% inhibition level. The A subunit and α chain preparations inhibited the binding of the radiolabeled B subunit to antitoxin sites. Conversely, B subunit inhibited the binding of radiolabeled A subunit and α chain to antitoxin. The γ chain did not show any reaction with antitoxin or cross-reaction with either whole toxin or its components. These results strongly suggest that the A subunit and the α chain contain antigenic determinant(s) that are common to the B subunit. The B subunit (β chain) and the α chain of cholera toxin may therefore contain region(s) of chemical similarity.
机译：描述了用于检测霍乱毒素及其组成多肽链的放射免疫测定方法。通过氯胺T法碘化霍乱毒素，A亚基，B亚基，α链和γ链。放射性标记并没有显着改变毒素或其组分的聚丙烯酰胺电泳迁移模式。此外，放射性标记的毒素，B亚基和α链制剂保留了与肠粘膜匀浆结合的基本能力。用放射性标记的毒素可检测到的最小抗毒素量为0.04抗毒素单位/ ml。用放射性标记的B亚基，A亚基和α链替代放射性标记的毒素会降低测试的敏感性。放射性标记的γ链不结合抗毒素制剂。竞争性抑制研究采用滴定的抗胆碱原血清和放射性标记的毒素或组分进行，结果表明，在90％的抑制水平下，可检测到的最低毒素浓度为7.0×10 μmol/ ml。 A亚基和α链制剂抑制了放射性标记的B亚基与抗毒素位点的结合。相反，B亚基抑制放射性标记的A亚基和α链与抗毒素的结合。 γ链与抗毒素没有反应，也没有与完整毒素或其成分发生交叉反应。这些结果强烈表明，A亚基和α链含有与B亚基相同的抗原决定簇。因此，霍乱毒素的B亚基（β链）和α链可能包含化学相似性区域。

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