水流动力学方法进行基因免疫制备高效价抗体  

Gene immunization for generation of high-level polyclonal antibodies by hydrodynamics-based procedure

作　　者：袁艳军[1] 王艳鸽[1] 刘峰涛[1] 马远方[1] 

机构地区：[1]河南大学医学院,免疫学研究所,河南省天然药物与免疫工程重点实验室,开封475004

出　　处：《现代免疫学》2009年第5期401-404,共4页Current Immunology

基　　金：河南省杰出人才创新基金资助项目

摘　　要：用水流动力学方法进行基因免疫,制备小鼠高效价多克隆抗体。采用水流动力学方法和肌肉注射pcDNA3.0/DR5免疫小鼠,分批在0、2、4、6周进行免疫,初次免疫8周后取鼠尾血用ELISA方法检测小鼠抗DR5抗体效价,用Westernblot、免疫组化染色法检测抗体的特异性。结果表明,ELISA结果显示末次免疫后水流动力学免疫组抗体滴度最高可达到为1∶102 400,肌肉注射免疫组抗体滴度最高可达1∶25 600。Western blot结果显示,小鼠抗DR5多克隆抗体识别Jurkat细胞的DR5蛋白条带。免疫组化结果表明,转染的DR5蛋白主要分布在食管癌细胞膜附近,与预期相符。用水流动力学的方法可以成功制备比肌肉注射更高效价的多克隆抗体。To prepare high-level mouse polyclonal antibody, mice were immunized with plasmid pcDNA3.0/DR5 for 4 times by intramuscular injections and the hydrodynamics-based procedure respectively （delivered on 0, 2, 4, 6 weeks） and its specificity was identified by Western blot assay and immunohistochemistry. As demonstrated by ELISA measurements, the antibody titer of group with hydrodynamics-based procedure and of group intramuscular injection could approach up to 1 ： 102 400 and 1 ： 25 600 respectively. The result of Western blot assay indicated that the mouse anti-DR5 polyclonal antibody could recognize the DR5 protein bands produced in Jurkat cells and the result of immunoehemistry showed that the transfected DR5 protein mainly distributed near the cellular membrane of esophageal cancer cells, as it was suspected. It is evident that high-titered polyclonal antibodies can be obtained by using the hydrodynamics procedure in comparison with those obtained by intramuscular injection.

关 键 词：水流动力学 肌肉注射 多克隆抗体 DR5 

分 类 号：R392-33[医药卫生—免疫学]