摘要
目的:制备单克隆抗体OX7(monoclonal antibody OX7,mAb OX7)及建立大鼠抗Thy1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,Protein A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2.06 mg/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OX7细胞注入小鼠腹腔并用Protein A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mAb OX7,该抗体可成功建立抗Thy1系膜增生性肾炎动物模型。
To prepare the OX7 monoclonal antibody （mAb OX7） and establish the anti - thyl mesangial proliferative nephritis model. Methods：Ascitic fluid was obtained from pristineprimed Balb/c mice intraperitoneally injected with OX7 - producing hyhridoma. Immunoglobulin G （IgG） of OX7 was further purifi - ed by protein A affinity column chromatography and used for the study. The purity d the antibody were identified by the sodium dodecyl sulfate polyacrylamide gel electropheresis（SDS - PAGE）. The purified mAb OX7 were intravenously administrated into wistar rats. Histophathologieal change of kidney sections were determined at day 0,3 and 7 days after intravenous injection. Results：No other straps but heavy chain and light chain of the purified antibody were found in the SDS - PAGE, and the antibody concentration was 2.06 mg/ml. At 3rd day, a large number of destroyed mesangial cells, dissolved mesangial matrix were observed. PAS staining showed that mesangial cell proliferation started at day 3, peaked at day 7, the time when proliferation is maximal in this model. Conclusion： We prepare high- valence,purified and specific the mAb OX7 （by injecting OX7 cells to mice with purifed ascites by Protein A affinity chromatography, which successfully establish the mesangial proliferative nephritis model.
出处
《中国中西医结合肾病杂志》
2009年第2期101-104,I0001,共5页
Chinese Journal of Integrated Traditional and Western Nephrology
基金
国家自然科学基金重点项目(No.30630033)
