用软琼脂糖培养与免疫吸附法制备鸡胚视网膜单克隆抗体  

Application of Soft Agarose Culture and Filter Immunoadsorption Assay in the Production of Monoclonal Antibodies against Embryonic Chick Retina

作　　者：李希斌[1] 刘正梅[1] T.Chen M.Nirenberg 

机构地区：[1]北京市神经外科研究所,北京100050 [2]美国国立卫生研究院生化遗传实验室

出　　处：《免疫学杂志》1991年第2期118-121,共4页Immunological Journal

摘　　要：细胞融合后,在软琼脂糖培养基上形成杂交瘤克隆,如将附有细胞或其他抗原的滤膜置于培养基表面,杂交瘤细胞分泌的抗体通过扩散与滤膜上的抗原结合,经与^(125)Ⅰ标记的第二抗体反应后,将滤膜进行放射自显影以显示阳性克隆的位置。用本法能快速大规模筛选特异性杂交瘤克隆,已用此法得到多株针对鸡胎视网膜的单克隆抗体。BALB/c mice were immunized with embryonic chick neural retina.After fusion hybridoma colonies were formed in the top layer of soft agarose culture medium. Milipore filters with cultured neural retina cells or isolated chick brain synaptosomes were placed onto the top of the agarose plates. During the incubation period monoclonal antibody molecules secreted py the hybridoma colonies bind specifically to the antigens on the filter. The filters were then incubated in the buffer containing 125I labeled goat anti-mouse IgG, IgA and IgM.After autoradiography the position of positive hybridoma colonies were indicated by the presence of dark spots on the X-ray film. This method is suitable for the generation of large number of specific hybridoma clones. Monoclonal antibodies reacting with different cellular or synaptical layers of chick neural retina has been produced with this method.

关 键 词：单克隆抗体 杂交瘤 鸡胚 视网膜 

分 类 号：R392－33[医药卫生—免疫学]