肺炎支原体种特异性单克隆抗体杂交瘤制备及鉴定

Preparation and identification of monoclonal antibody specific for Mycoplasma pneumoniae

目的 制备抗肺炎支原体种特异性单克隆抗体。方法 以纯化的肺炎支原体膜蛋白抗原 (相对分子质量为43× 10 3)加福氏免疫佐剂免疫 8周龄雌性BALB c小鼠 ,取小鼠脾细胞与骨髓瘤细胞SP2 0 Ag14融合 ,以酶标记免疫吸附测定法 (ELISA)筛选和鉴定所制备的单克隆抗体。结果 共筛出 3株分泌抗肺炎支原体种特异单克隆抗体杂交瘤 ,经免疫鉴定其分泌的免疫球蛋白类型为IgG1、IgG2a及IgM ,与肺炎支原体有很强的亲和力 ,与种属相近的其它 6株支原体没有交叉反应。结论 本次筛选制备的单克隆抗体杂交瘤能分泌高浓度的肺炎支原体单克隆抗体 ,并且有很强的种特异性。

Objective To prepare anti-Mycoplasma pneumoniae monoclonal antibody. Methods 8-week female BALB/c mouse was immunized by the purified membrane protein antigen with immunoadjuvant, its relative molecular weight was 43×103. After, fusion between spleen cells from a mouse and SP2/0-Ag14 mouse myeloma cells was done. Monoclonal antibodies were screened in indirect enzyme-labeled immunosorbent assay (ELISA). Results Three hybridomas producing monoclonal antibodies specific for Mycoplasma pneumoniae were obtained. The type of immunoglobulin which three hybridomas produced were identified for IgG1, IgG2a and IgM by immunology method. None of the three monoclonal antibodies was implicated in immunological cross-reactivity with other isolates of Mycoplasma. Conclusion The three monoclonal antibodies were specific towards Mycoplasma pneumoniae.