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| 抗酶单克隆抗体制备中检测方法的探讨 | |
| 引用本文： | 欧阳斌,刘燕,陈敬,史良如. 抗酶单克隆抗体制备中检测方法的探讨[J]. 中国生物制品学杂志, 1990, 0(3) |
| 作者姓名： | 欧阳斌 刘燕 陈敬 史良如 |
| 作者单位： | 卫生部武汉生物制品研究所，卫生部武汉生物制品研究所，卫生部武汉生物制品研究所，卫生部武汉生物制品研究所 武昌 430060，武昌 430060，武昌 430060，武昌 430060 |
| 摘 要： | 本文比较了反向间接酶联免疫吸附法和传统的间接酶联免疫吸附法。发现前者更适合作为抗酶单克隆抗体制备中的筛选方法。并对试验中的各种反应条件,包括羊抗鼠IgG包板、待检单抗和酶反应的最适浓度及底物缓冲液的选择进行了探索,最后用该法筛选5次融合后培养的杂交瘤上清,对其结果进行了分析和评价。 |
| 关 键 词： | 硷性磷酸酶(AP) 抗硷性磷酸酶(AAP) 反向间接酶联免疫吸附法 单克隆抗体(McAb) |
Studies on the Screening Methods of Preparing Monoclonal Antibodies Recognized Enzymes |
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| Ouyang Bin et al Wuhan Institute of Biological Products,Wuhan. Studies on the Screening Methods of Preparing Monoclonal Antibodies Recognized Enzymes[J]. Chinese Journal of Bilogicals, 1990, 0(3) | |
| Authors: | Ouyang Bin et al Wuhan Institute of Biological Products Wuhan |
| Abstract: | Reverse indirect enzyme linked immunoadsorption and indirect enzyme linked immunoadsorption for detection of monoclonal antibodies recognized enzymes were compared and analysed in this paper. It was demonstrated that the former method was a good procedure for screening. The various conditions of the reaction were also investigated. The results indicated that sheep anti-mouse IgG coated to plate with concentration of 500μg/ml and 250μg/ml respectively and alkaline phosphatase reacted with 2mg/ml (Sigma I powder) were optimal. It wasn't necessary to dilute the hybridoma cell culture supernatant when screening the monoclonal antibodies. Of three substrate buffers, pH9.8 , 10% diethanolamine was the best one. Finally, we have used the reverse indirect enzyme linked immunoadsorption to screen hybridoma supernatant cultured after fusion and obtained 7 cell lines of hybridomas secreting anti-alkaline phosphatase monoclonal antibodies. |
| Keywords: | Alkaline phosphatase Anti-alkaline phosphatase Reverse indirect enzyme linked immunoadsorption Monoclonal antibody |
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