单克隆抗体OX7的制备及大鼠抗Thy1系膜增生性肾炎模型的建立

目的：制备单克隆抗体OX7（monoclonal antibody OX7,mAbOX7）7i建立大鼠抗Tbyl系膜增生性肾炎模型。方法：弗氏不完全佐剂预免疫Balb／c小鼠,腹腔注射OX7细胞,ProteinA亲和层析法纯化腹水,用SDS—PAGE鉴定纯化后抗体的纯度。将所得抗体尾静脉注射Wistar大鼠,PAS染色观察正常对照组注射前及注射后第3天、第7天大鼠肾脏病理组织学改变。结果：SDS—PAGE电泳显示,纯化的抗体有IgG的轻链和重链两条带,无其他杂带,抗体浓度为2．06mg／ml。PAS染色显示正常对照组肾小球结构正常,毛细血管袢开放良好;模型组第3天开始出现大量系膜细胞破坏,系膜基质溶解,肾小球毛细血管扩张;第7天系膜细胞重度增生。结论：通过将0X7细胞注入小鼠腹腔并用ProteinA亲和层析法纯化腹水,可成功制备高效价、高纯度及特异性强的mAbOX7,该抗体可成功建立抗Thyl系膜增生性肾炎动物模型。

Preparation of Monoclonal Antibody OX7 and Establishment of Anti-thy 1 Mesangial Proliferative Nephritis Model in Rats

Objective:To prepare the OX7 monoclonal antibody(mAb OX7) and establish the anti-thy1 mesangial proliferative nephritis model.Methods:Ascitic fluid was obtained from pristine-primed Balb/c mice intraperitoneally injected with OX7-producing hybridoma.Immunoglobulin G(IgG) of OX7 was further purifi-ed by protein A affinity column chromatography and used for the study.The purity of the antibody were identified by the sodium dodecyl sulfate polyacrylamide gel electropheresis(SDS-PAGE).The purified mAb OX7 were ...