Chang L S, Lin J, Chang C C
Department of Biochemistry, Kaohsiung Medical College, Taiwan, ROC.
Biochem Mol Biol Int. 1995 Apr;35(4):733-8.
Antibodies which specifically reacted with reduced and S-carboxy-methylated(RCM) cobrotoxin were purified from anticobrotoxin antisera and anti-RCM-cobrotoxin antisera, respectively. Results using a competitive immunoassay revealed that the antibodies from anti-RCM-cobrotoxin antisera had a greater affinity for RCM-cobrotoxin than for cobrotoxin. Whereas the reactivity of the antibodies from the anticobrotoxin antisera toward cobrotoxin and RCM-cobrotoxin had a reversed order of binding. In contrast to observations made with S. aureus V8 protease-digest hydrolysates, the antigenic structures of RCM-cobrotoxin recognized by antibodies from anti-RCM-cobrotoxin antisera were notably affected following hydrolysis of RCM-cobrotoxin with chymotrypsin. Moreover, the chymotryptic hydrolysates showed a comparable reactivity as RCM-cobrotoxin toward the antibodies purified from anticobrotoxin antibodies, but a decrease in antigenicity with the V8 protease hydrolysates. These results reveal that the repertoire of antibodies against the unfolded cobrotoxin are not the same in anticobrotoxin and anti-RCM-cobrotoxin antisera. Moreover, it suggests that the repertoire of antibodies from the different sources against the same antigen can be differentiated by measurement of their reactivities with the proteolytic hydrolysates of the antigen.
分别从抗眼镜蛇毒素抗血清和抗RCM - 眼镜蛇毒素抗血清中纯化出与还原型和S - 羧甲基化（RCM）眼镜蛇毒素特异性反应的抗体。竞争免疫分析结果显示，抗RCM - 眼镜蛇毒素抗血清中的抗体对RCM - 眼镜蛇毒素的亲和力比对眼镜蛇毒素的亲和力更高。而抗眼镜蛇毒素抗血清中的抗体对眼镜蛇毒素和RCM - 眼镜蛇毒素的反应性在结合顺序上则相反。与用金黄色葡萄球菌V8蛋白酶消化水解产物的观察结果不同，用胰凝乳蛋白酶水解RCM - 眼镜蛇毒素后，抗RCM - 眼镜蛇毒素抗血清中的抗体所识别的RCM - 眼镜蛇毒素的抗原结构受到显著影响。此外，胰凝乳蛋白酶水解产物对从抗眼镜蛇毒素抗体中纯化的抗体显示出与RCM - 眼镜蛇毒素相当的反应性，但V8蛋白酶水解产物的抗原性降低。这些结果表明，抗眼镜蛇毒素抗血清和抗RCM - 眼镜蛇毒素抗血清中针对未折叠眼镜蛇毒素的抗体库是不同的。此外，这表明可以通过测量来自不同来源的抗体与抗原的蛋白水解产物的反应性来区分它们针对同一抗原的抗体库。
