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金黄色葡萄球菌肠毒素B(SEB)属于热毒素超抗原家族。它通过激活T细胞,使T细胞大量增殖并释放大量炎症细胞因子,从而导致中毒性休克综合征及死亡。为了筛选获得抗SEB的中和保护性单克隆抗体,我们首先克隆了SEB基因,表达纯化获得GST-SEB融合蛋白;将之免疫BALB/c小鼠后,利用杂交瘤技术结合ELISA检测,筛选获得10株抗SEB单克隆抗体杂交瘤细胞株;ELISA和Western blot鉴定其分泌的抗体能与SEB特异结合;抗体亚型检测均为IgG1/k;体外实验证实,10株抗体中有6株(1A5、4A3、3E2、3C1、1D1、2G9)能够有效阻断SEB诱导的人外周血单核细胞(PBMC)的激活,具有中和保护作用。本研究为进一步获得抗SEB的人源化单克隆中和性抗体奠定了基础。
Staphylococcal enterotoxin B (SEB) belongs to the family of thermo toxin superantigens. It causes toxic shock syndrome and death by activating T cells, proliferating T cells in large numbers, and releasing large quantities of inflammatory cytokines. In order to obtain the neutralizing protective monoclonal antibody against SEB, we first cloned the SEB gene and expressed and purified the GST-SEB fusion protein. After immunizing the BALB / c mice, we obtained the SEB gene by hybridoma assay and ELISA 10 strains of monoclonal antibodies against SEB hybridoma cell lines; ELISA and Western blot identification of the secretion of antibodies with SEB specific binding; antibody subtypes were detected IgG1 / k; in vitro experiments confirmed that 10 out of antibodies in 6 (1A5 , 4A3, 3E2, 3C1, 1D1, 2G9) can effectively block the SEB-induced activation of human peripheral blood mononuclear cells (PBMC), with a neutralizing protective effect. This study laid the foundation for further obtaining humanized monoclonal neutralizing antibodies against SEB.