质粒DNA单次脾内注射制备单克隆抗体  

Preparation of Monoclonal Antibody Based on Single Intrasplenic Immunization of Plasmid DNA

作　　者：郑泓[1] 韩双艳[1] 林影[1] 王小宁[1] 

机构地区：[1]华南理工大学生物科学与工程学院,广州510640

出　　处：《生物工程学报》2007年第4期710-714,共5页Chinese Journal of Biotechnology

基　　金：国家科技攻关计划项目(No2004BA711A20)~~

摘　　要：探索一种新的快捷有效DNA免疫制备单克隆抗体的方法,辅助实现构建高通量无蛋白纯化体系单克隆抗体制备和筛选。分别通过“重叠PCR”和“无模板PCR”在pVAX1真核载体中分别引入IL-2信号肽、IgG kappa链信号肽构建分泌型真核表达载体,将代表抗原基因的profilin1基因克隆到经改造带有信号肽基因的表达载体上,构建重组质粒pVAX-IL2-prof1和pVAX-Igκ-prof1,单次脾内注射重组质粒DNA免疫BALB/c小鼠。经过细胞融合、ELISA筛选,获得两株抗profilin1的单克隆抗体。单抗亚型分别为IgM和IgG3。单次脾内质粒DNA免疫便捷有效,是制备单克隆抗体的有效方法。To set up a new rapid and efficient way for the preparation of specific monoclonal antibodies （MAbs） with plasmid DNA immunization. Methods The fusion gene of IL-2 signal peptide and profilin 1 by ＇overlapping PCR＇ was obtained and inserted into the vector pVAX1 to construct the recombinant plasmid pVAX-IL2-profl. Another fusion gene of IgG kappa chain signal peptide and profilin 1 by ＇ no template PCR＇ was obtained and inserted into the vector pVAX1 to construct the recombinant plasmid pVAX-Igx-profl. BALB/c mice were single intrasplenie immunized with plasmid DNA. Results After cell fusion and hybridomas screening by indirect ELISA, we charactered two mAbs （ 1D8A2B4 and 4B12A5E3） against profilin 1. The MAbs isotype were determined as IgM and IgG3. Conclusion A single intrasplenic plasmid DNA immunization is rapid and efficient and can be used as a helpful tool for the production of specific mAb.

关 键 词：脾内DNA免疫 单克隆抗体 profilin 1 

分 类 号：Q813[生物学—生物工程]