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分泌抗黄曲霉毒素M1抗体的大鼠-小鼠杂交瘤的建立及其单克隆抗体的特性

The establishment of rat-mouse hybridomas secreting anti-aflatoxin M1 antibodies and the properties of their monoclonal antibodies.

作者信息

Ji Y, Lin G, Ye M

机构信息

Shanghai Institute of Cell Biology, Chinese Academy of Sciences.

出版信息

Chin J Biotechnol. 1992;8(2):113-21.

PMID:1297440
Abstract

After a comparison of anti-AFM1-BSA antibody responses between rat and mouse, the spleen cells of rat with stronger responses were chosen as parent cells for fusing with mouse myeloma cells P3X63-Ag8.653. Through HAT medium selection, RIA screening and cloning, five well growing rat-mouse hybridoma clones were obtained that could secret anti-AFM1 antibodies stably. The results from ELISA and competitive binding RIA further proved that the 5 McAbs are direct against AFM1, with significant cross reaction to its derivative, AFB1. The average affinity constant of the 5 McAbs is 10(9)-10(11) l/M. It signifies that these monoclonals have potential application value for the construction of AF detection kit.

摘要

比较大鼠和小鼠抗AFM1-BSA抗体反应后,选择反应较强的大鼠脾细胞作为亲本细胞,与小鼠骨髓瘤细胞P3X63-Ag8.653进行融合。通过HAT培养基筛选、RIA筛选和克隆,获得了5个生长良好的大鼠-小鼠杂交瘤克隆,它们能够稳定分泌抗AFM1抗体。ELISA和竞争性结合RIA结果进一步证明,这5株单克隆抗体直接针对AFM1,对其衍生物AFB1有明显交叉反应。5株单克隆抗体的平均亲和常数为10(9)-10(11) l/M。这表明这些单克隆抗体在构建AF检测试剂盒方面具有潜在的应用价值。

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