Flatt P R, Swanston-Flatt S K
Endocrinol Exp. 1981;15(1):3-16.
Rabbits and guinea pigs were immunized monthly with a glucagon-bovine plasma albumin conjugate prepared at a molar ratio of 12: 1 using 1-ethyl-3(3-dimethyl-aminopropyl)-carbodiimide as the coupling agent. The animals were bled regularly two weeks after inoculation and the resulting glucagon antisera analyzed for titre, affinity and specificity. Immunization of rabbits resulted in the production of large amounts of high affinity glucagon antibodies which in two out of five animals reacted between 3 and 7% with GLI (glucagon-like immunoreactivity). Antibody titre and affinity rose rapidly in the majority of rabbits attaining near maximal values by 10 weeks. In comparison, the conjugate displayed enhanced immunogenicity in the guinea pig, and antibody titre and affinity which were already several fold higher at 10 weeks continued to rise progressively during the 26 week period of immunization. Antisera raised in guinea pigs reacted particularly strongly with GLI, and antibodies derived from one particular animal yielded practically identical dilution curves with glucagon and GLI. The present study clearly demonstrates the potential of the glucagon-carbodiimide-albumin conjugate for the production of valuable glucagon antisera in rabbits and guinea pigs. Such antisera are eminently suitable for radioimmunoassay and metabolic studies. Furthermore, it is concluded that the guinea pig is a particularly useful species for the rapid production of very high quality cross-reacting glucagon antisera.
使用1-乙基-3-(3-二甲基氨基丙基)碳二亚胺作为偶联剂，以12:1的摩尔比制备胰高血糖素-牛血清白蛋白偶联物，每月对兔子和豚鼠进行免疫。接种两周后定期采集动物血液，分析所得胰高血糖素抗血清的效价、亲和力和特异性。对兔子进行免疫导致产生大量高亲和力的胰高血糖素抗体，五只动物中有两只与胰高血糖素样免疫反应性(GLI)的反应率在3%至7%之间。大多数兔子的抗体效价和亲和力迅速上升，到10周时达到接近最大值。相比之下，该偶联物在豚鼠中显示出增强的免疫原性，10周时已经高出几倍的抗体效价和亲和力在26周的免疫期间继续逐渐上升。豚鼠产生的抗血清与GLI反应特别强烈，来自一只特定动物的抗体与胰高血糖素和GLI产生几乎相同的稀释曲线。本研究清楚地证明了胰高血糖素-碳二亚胺-白蛋白偶联物在兔子和豚鼠中产生有价值的胰高血糖素抗血清的潜力。这种抗血清非常适合放射免疫分析和代谢研究。此外，得出的结论是，豚鼠是快速产生高质量交叉反应性胰高血糖素抗血清的特别有用的物种。
