摘要
目的以免疫亲和层析方法纯化重组人促红细胞生成素(rhEPO)并免疫新西兰家兔制备多克隆抗体,用辣根过氧化物酶标记抗体,为进一步建立ELISA检测体系做准备。方法购买rhEPO,用淋巴结内注射法制备多克隆抗体,EPO抗体与葡聚糖凝胶偶联,制成抗体亲和层析柱,以rhEPO作为抗原上样纯化。抗体先盐析粗提纯再过SPA亲和层析柱纯化。用高碘酸钠法制备酶标抗体。结果亲和层析纯化的抗原、抗体纯度较高,蛋白质含量测定分别为0.560、.41 mg/ml;酶结合率为0.578。结论免疫亲和层析法纯化rhEPO,制备的多克隆抗体及酶标记抗体的纯度好,可以满足下一步组建试剂盒的需求。
Objective Purified the rhEPO using the immunoaffinity chromatography method,and immunized New Zealand rabbits to produce the EPO polyclonalantibody.Labeled the antibody with HRP,to prepare for establishing ELISA system.Methods Purchased the rhEPO,coupled the EPO antibody with sephadex to make the affinity column.Then,purify the rhEPO as the sample.With regard to the antibody,it should be primarily purified by the salting-out,and then purified with the SPA affinity column.Prepared the enzyme labelled antibody by the sodium metaperiodate method.Results The EPO antibody and antigen were highly purified,the protein contents were 0.56 and 0.41 mg/ml.the enzyme biding rate was 0.578.Conclusion Purified the rhEPO and enzyme labeled antibody using the immunoaffinity chromatography method and the they were highly purified,so it met the need of making the ELISA syystem.
出处
《中国实验诊断学》
北大核心
2011年第10期1611-1614,共4页
Chinese Journal of Laboratory Diagnosis
基金
天津市科技型中小企业技术创新专项资金项目(08ZXCXSH03300)
