抗腐败西瓦菌单克隆抗体杂交瘤细胞系的建立及初步鉴定

目的 制备和鉴定抗腐败西瓦菌单克隆抗体(mAbs),为建立重要海洋细菌快速ELISA检测试剂盒奠定基础.方法 用灭活腐败西瓦菌免疫小鼠,利用杂交瘤细胞融合技术,制备出抗腐败西瓦菌杂交瘤细胞株,用Giemsa染色对其进行染色体鉴定,并用间接ELISA方法筛选、鉴定其与腐败西瓦菌及其他重要海洋细菌的交叉反应性和效价.结果 共获得4株抗腐败西瓦菌的杂交瘤细胞株(02D6、02H12、03G4、03A7),均符合杂交瘤细胞染色体特点,其培养上清效价为10-1～10-2,具有良好的特异性和免疫反应性.结论 研究制备、筛选出抗腐败西瓦菌的特异性抗体,有助于进一步建立重要海洋细菌快速检测试剂盒.

Preparation and initial characterization of monoclonal antibody against Shewanella putrefaciens

OBJECTIVE To prepare high-performance and specific monoclonal antibody(mAbs) against Shewanella putrefaciens so as to establish rapid ELISA kit of the important marine bacteria.METHODS BALB/c mice were immunized with S.putrefaciens,and hybridoma cells against S.putrefaciens were produced by cell-fusion technology.The chromatosomes of hybridoma cells were identified by Giemsa,the potency and cross-reactivity against S.putrefaciens as well as other important marine bacteria were screened by ELISA.RESULTS We obtained four strains of hybridoma against S.putrefaciens(02D6,02H12,03G4 and 03A7),all of which had the characteristics of hybridoma cells.The titer ranges of the cultivation supernatant of hybridoma was 10-1-10-2.The hybridoma had favourable specifity and immunoreactivity.CONCLUSION Specific antibodies against S.putrefaciens are produced in this study,which might also be useful for establishing rapid-detection kit of the important marine bacteria.