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目的制备并鉴定2型猪链球菌重组Sao-M蛋白的单克隆抗体(Monoclonal Antibody,McAb),为建立免疫学快速检测方法及Sao蛋白的功能研究奠定基础。方法用重组Sao-M蛋白免疫BALB/c小鼠,利用细胞融合技术建立稳定分泌抗Sao-M蛋白的杂交瘤细胞株,制备抗Sao单克隆抗体,采用IgG亚类鉴定试剂盒鉴定单克隆抗体的IgG亚型,采用间接ELISA法测定抗体效价。将05ZYH33与单抗共孵育,检测单克隆抗体与细菌表面具有天然构象Sao蛋白的结合能力。结果筛选到4株能持久、稳定分泌抗Sao蛋白单克隆抗体的杂交瘤细胞株(分别命名为1G5、1B10、2A10、3A6),均为IgG1型,ELISA测定腹水抗体效价>1︰102 400;4株单克隆抗体均能与05ZYH33表面天然构象的Sao蛋白结合,对细菌形态产生显著影响。结论成功制备高效价的抗Sao-M单克隆抗体,为建立快速检测猪链球菌感染的免疫学方法及Sao蛋白的功能研究奠定了基础。
Objective To prepare and identify Monoclonal Antibody (McAb) against recombinant Sao-M protein of Streptococcus suis type 2, and to lay the foundation for the establishment of a rapid immunological assay and the functional study of Sao protein. Methods BALB / c mice were immunized with recombinant Sao-M protein, and the hybridoma cell line secreting anti-Sao-M protein was established by cell fusion technique to prepare anti-Sao monoclonal antibody. The McAbs were identified by IgG subclass identification kit Of IgG subclasses were measured by indirect ELISA antibody titer. 05ZYH33 co-incubated with monoclonal antibody to detect monoclonal antibody and bacterial surface with the natural conformation Sao protein binding capacity. RESULTS: Four hybridoma cell lines secreting anti-Sao protein monoclonal antibodies (1G5, 1B10, 2A10 and 3A6, respectively) were screened out for long-term and stable IgG1 antibody titer> 1: 102 400 ; All four monoclonal antibodies could bind to Sao protein of natural conformation on the surface of 05ZYH33, which had a significant effect on the bacterial morphology. Conclusion The successful preparation of high titer anti-Sao-M monoclonal antibody laid the foundation for the establishment of an immunological method for the rapid detection of S. suis infection and the functional study of Sao protein.