福氏志贺氏菌2a单克隆抗体的制备及ELISA方法的建立

摘要

为制备特异性和灵敏度较高的抗福氏志贺氏菌2a单克隆抗体,并建立双抗夹心酶联免疫吸附检测法,用于检测生鲜食品中的福氏志贺氏菌2a.本研究利用自制的福氏志贺氏菌2a抗原免疫BALB/c小鼠,并用杂交瘤技术进行细胞融合,经筛选克隆后,获得3株分泌抗福氏志贺氏菌2a单克隆抗体稳定的杂交瘤细胞,分别命名为5C12、1B5和6A11.3株杂交瘤细胞注射进小鼠腹腔后,诱导产生的腹水中抗体的效价为 1∶2.048×105~1∶1.024×105,免疫球蛋白亚型均为IgG.3株抗体识别抗原表位的最佳配对为5C12和6A11,由此建立双抗夹心ELISA体系,测得其灵敏度为1 000 CFU/g,且具有良好的特异性.%Preparation of specific and highly sensitive Shigella flexneri 2a monoclonal antibodies.The enzyme-linked immunosorbent assay (ELISA) was developed for the detection of Shigella flexneri 2a in food.In this study, BALB/c mice were immunized with Shigella flexneri 2a antigen.Hybridization techniques were used for cell fusion.After screening and cloning, three hybridoma cells lines, 5C12, 1B5 and 6A11, were obtained secreting monoclonal antibodies against S.flexneri 2a stably.Three hybridoma cells were named.The hybridoma cells was injected into the abdominal cavity of mice, and the induced antibodies titers were 1∶1.024×105、1∶1.024×105 and 1∶2.048×105.Their immune globulin subtypes were IgG.The best pairing of three antibodies was 5C12 and 6A11.The sandwich ELISA system established afterwards was good in sensitivity and specificity.