欲建立人類胰島素的臨床分析前,為提高其靈敏度、精確性,對於抗體的來源採用融合瘤技術來製備單源抗體。此過程藉少量豬胰島素直接對bal b/c小白鼠脾臟免疫四天後,即與NS- 1骨癌細胞融合,經HAT培養液選別,單殖化、酵素免疫分析法篩選,得到一株融合瘤,它可分泌Ig M單源抗體,所含的輕鏈為k鍵,其等電點為9.6 ± 0.1,力價為2,048,與蛋白質A無結合能力。對人的胰島素有作用,初步簡單試驗可測至8 μg/ml的胰島素濃度。 Monoclonal antibody against porcine insulin, prepared by hybridoma cell culture of human insulin clinical analysis technique, was used to increase sensitivity and precision.
Ba1b/c mice were intrasplenically inoculated with small amount of porcine insulin for 4 days, then their spleen cells were selected with HAT medium, cloning and screened by enzyme-immunoassay. After proper propagation, an IgM form monoclonal antibody with kappa light Chain, pI value of 9.6 ± 0.1, titer of 2048 was obtained. It bound to human insulin but not to protein A. Preliminary results show as low as 8 μg/ml of human insulin can be assayed by double sandwich ELISA method using a sorbent of HRP-linked antibody obtained in this study.