B19单倍型鸡BNK蛋白单克隆抗体的制备  

Establishment of Cell Line Expressing Chicken B19 BNK Protein Stably and Production of BNK19-Specific Monoclonal Antibody

作　　者：吴少莲 彭蔚宇[1] 王兴童 文辉强[1] 廉传江[1] 陈洪岩[1] 韩凌霞[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所实验动物与比较医学团队/兽医生物技术国家重点实验室,哈尔滨150001

出　　处：《实验动物科学》2015年第3期1-5,共5页Laboratory Animal Science

基　　金：国家自然科学基金(No:31101800)

摘　　要：目的体外构建稳定表达鸡自然杀伤细胞(Natural killer,NK)BNK蛋白的细胞系,制备BNK蛋白的单克隆抗体,为鸡BNK蛋白及NK细胞的研究提供便利。方法利用BNK19原核表达产物免疫BALB/c小鼠,体外稳定表达疫病敏感性B19单倍型鸡BNK蛋白(BNK19)的真核细胞系筛选单克隆抗体。结果用稳定表达BNK蛋白的真核细胞系筛选到2株BNK19特异性单克隆抗体。结论获得的单克隆抗体经鉴定具有免疫学活性,为鸡BNK蛋白及NK细胞的作用机制研究提供了条件。Objective To product BNK19-specific monoclonal antibody using the B19 haplotype chicken BNK protein expressed by a stable cell line. Method B-NK19 gene was amplified from B19 haplotype chicken and then cloned into the p IRES vector to construct recombinant plasmid of p IRES-BNK19. The recombinant plasmid was transfected into CHO cell and identified by RT-PCR and indirect immunofluorescent assay( IFA). BALB / c mice were immunized using the BNK protein expressed by the recombinant E. coli,and the monoclonal antibody was selected from the CHO-BNK19 cell line. Result The CHO-BNK19 cell line expressing BNK protein stably was identified and two specific strains of monoclonal antibody were obtained using IFA. Conclusion The study is expected to be useful for researching chicken BNK protein and NK cells.

关 键 词：B19单倍型鸡 BNK 单克隆抗体 

分 类 号：S852.4[农业科学—基础兽医学]