B19 单倍型鸡 BNK 蛋白单克隆抗体的制备

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。

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

Abstract: 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 pIＲES vector to construct recombinant plasmid of pIＲES-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．