Court G
Pathol Biol (Paris). 1975 Dec;23(10):859-62.
Three factors which affect the outcome of any immunisation schedule designed to produce antisera for radio-immunoassay, the antigen, the method of immunisation and the choice of animal are considered. Several factors concerning the nature of the antigen are dealt with, for example, the molecular size and immunogenicity of the antigen. It is noted that the larger polypeptide and proteins are sufficiently immunogenic to elicit a useful antibody response alone and that whilst substances with molecular weights of less than 2000 may produce a response alone they will probably produce a better one if they are conjugated (chemically coupled) to a much larger molecule. With the low molecular weight substances such as the thyroxine, steroid hormones, and drugs which are of much clinical interest to many people conjugation is essential. The availability and purity of the immunogen are considered, especially in cases where a minute amount of material has been produced following a long and tedious purification procedure. The necessity in such instances that workers are aware of the smallest amount of material that will produce a useful antibody response and also of the most effective method of immunisation is stressed. The method of immunisation is discussed including a consideration of the use of adjuvant and the route and timing of injections. It is noted that antisera showing the relevant properties for radio-immunoassay are rarely produced without emulsification of the immunogen in Freund's adjuvant although this is not an absolute requirement for antibody production. Data are presented comparing the intramuscular and multiple intradermal routes of injection. The results, however, fail to demonstrate any major advantage for either method although the latter may be more economical, producing high titre antisera with relatively small amounts of immunogen. Because of their convenience rabbits are generally the first choice of animal for raising antisera for radioimmunoassay although guinea pigs, chickens and sheep have been used successfully in many cases.
本文考虑了影响任何旨在生产用于放射免疫分析抗血清的免疫方案结果的三个因素，即抗原、免疫方法和动物的选择。讨论了与抗原性质相关的几个因素，例如抗原的分子大小和免疫原性。需要注意的是，较大的多肽和蛋白质具有足够的免疫原性，能够单独引发有效的抗体反应；而分子量小于2000的物质虽然可能单独产生反应，但如果与大得多的分子偶联（化学结合），可能会产生更好的反应。对于许多人非常感兴趣的低分子量物质，如甲状腺素、类固醇激素和药物，偶联是必不可少的。还考虑了免疫原的可得性和纯度，特别是在经过漫长而繁琐的纯化过程后仅产生微量物质的情况下。强调了在这种情况下，工作人员必须了解产生有效抗体反应所需的最小物质量以及最有效的免疫方法。文中讨论了免疫方法，包括对佐剂的使用、注射途径和时间的考虑。需要注意的是，尽管弗氏佐剂乳化免疫原并非产生抗体的绝对必要条件，但在不进行这种乳化的情况下，很少能产生具有放射免疫分析相关特性的抗血清。文中给出了比较肌肉注射和多次皮内注射途径的数据。然而，结果未能证明这两种方法有任何主要优势，尽管后者可能更经济，能用相对少量的免疫原产生高滴度抗血清。由于其便利性，兔子通常是用于制备放射免疫分析抗血清的首选动物，尽管豚鼠、鸡和羊在许多情况下也已成功使用。
