Allauzen S, Salhi S L, Piechaczyk M, Bastide M, Pau B, Bouanani M
CNRS UMR 9921, UFR Pharmacie, Montpellier, France.
Immunol Lett. 1994 Apr;40(1):1-6. doi: 10.1016/0165-2478(94)90197-x.
A new immunization protocol has been developed to obtain specific monoclonal antibodies (mAb) directed against the soluble form of human insulin. Various protocols differing on the basis of the nature of immunogen, the number of injections and the route of administration of the antigen were compared. Mice with the highest anti-insulin titers were selected for cell fusion. The results showed that the immunization protocol involving 2 injections of insulin followed by a boost 2 months later mainly stimulated B lymphocytes secreting IgM mAb directed against immobilized insulin. Immunization with 2 injections of human proinsulin followed by 2 injections of a human insulin-bovine serum albumin conjugate and finally with a booster injection of this conjugate on each of the last 4 days preceding fusion was necessary to obtain a high percentage of hybridomas secreting specific IgG mAb able to recognize immobilized insulin (indirect ELISA) as well as iodinated insulin (liquid-phase radioimmunoassay).
已开发出一种新的免疫方案，以获得针对人胰岛素可溶性形式的特异性单克隆抗体（mAb）。比较了基于免疫原性质、注射次数和抗原给药途径不同的各种方案。选择抗胰岛素滴度最高的小鼠进行细胞融合。结果表明，免疫方案为注射2次胰岛素，2个月后加强免疫，主要刺激分泌针对固定化胰岛素的IgM mAb的B淋巴细胞。为了获得高比例的分泌特异性IgG mAb的杂交瘤，这些杂交瘤能够识别固定化胰岛素（间接ELISA）以及碘化胰岛素（液相放射免疫测定），需要先注射2次人胰岛素原，然后注射2次人胰岛素-牛血清白蛋白偶联物，最后在融合前的最后4天每天注射一次该偶联物作为加强免疫。
