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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
  • ISSN号:1009-587X
  • 期刊名称:《中国中西医结合肾病杂志》
  • 时间:0
  • 分类:R-332[医药卫生] R692.3[医药卫生—泌尿科学;医药卫生—临床医学;医药卫生—外科学]
  • 作者机构:[1]解放军总医院肾病科,全军肾脏病研究所暨重点实验室,北京100853, [2]哈尔滨医科大学附属第一医院肾内科,哈尔滨150001
  • 相关基金:本课题为国家自然科学基金重点项目(No.30630033)
中文摘要:

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。

英文摘要:

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.

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期刊信息
  • 《中国中西医结合肾病杂志》
  • 中国科技核心期刊
  • 主管单位:中国科学技术协会
  • 主办单位:中国中西医结合学会
  • 主编:
  • 地址:山西太原解放路162号(太原市06079信箱)
  • 邮编:030006
  • 邮箱:sx7965258@126.com;yhkj@public.ty.sx.cn
  • 电话:0351-7965258
  • 国际标准刊号:ISSN:1009-587X
  • 国内统一刊号:ISSN:14-1277/R
  • 邮发代号:22-26
  • 获奖情况:
  • 国内外数据库收录:
  • 中国中国科技核心期刊
  • 被引量:21630