胰島素酵素免疫分析中單源抗體的製備研究

CNU IR > Chna Nan Annual Bulletin > Vol.12 (1986) > Item 310902800/25047

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Title:	胰島素酵素免疫分析中單源抗體的製備研究 Studies on the Preparation of Monoclonal Antibody for Human Insulin Enzyme Immunoassay
Authors:	劉雅麗熊光濱
Contributors:	應用化學科工業技術研究院化學工業研究所
Date:	1986
Issue Date:	2012-02-24 16:02:19 (UTC+8)
Abstract:	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.
Relation:	嘉南學報 12：p.47-57
Appears in Collections:	[Chna Nan Annual Bulletin] Vol.12 (1986) [Dept. of Food & Drug Industry and Inspective Technology] Periodical Articles

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