金黄色葡萄球菌肠毒素单克隆抗体制备与应用
Production and preliminary application of anti-staphylococcal enterotoxinB monoclonal antibodies
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摘要:目的 制备抗B型金黄色葡萄球菌肠毒素单克隆抗体。方法 纯化B型金黄色葡萄球菌肠毒素(SEB)免疫BALB/C小鼠的脾细胞与SO2/0骨髓瘤细胞融合, 经筛选及克隆化, 共获得6株能稳定分泌抗B型金黄色葡萄球菌肠毒素单克隆抗体的杂交瘤细胞株并进行了鉴定。结果 6株单克隆抗体除两株属于IgG2A外, 其余均属于IGG1亚类, 其培养上清和腹水的滴度分别为1256~1512和112800~125600, 6株单克隆抗体均不与SEA发生交叉反应, 仅有2株与SEC1有弱交叉反应.WESTERNBLOT实验显示出一条特异性带, 位于分子量28.5KDA处, 证明了其特异性。结论 成功获得抗B型金黄色葡萄球菌肠毒素单克隆抗体, 为检测SEB打下了基础。Abstract:Objective To prepare Anti-staphylococcal EnterotoxinB(SEB)Moncolonal antibodies.Methods Hybridomas secreting Anti-SEB Monoclonal Antibodies were developed by fusion of SP2/0 myeloma cells and splenocytes from BALB/C mice immunized with purified SEB.Six positive clones Anti-SEB Monoclonal Antibodies were obtained by repeated screening and cloningsResults 6 strains belonged to Ig Glsubclass except two strains(IgG2-a)and their titers in culture supernatant and ascitic fluid by ELISA were between 1:256~1:512 and 1:12 800~1:25 600 respectively, It was revealed that 2 strains had very weak cross-reaction to SEC1, All Monoclonal Antibodies recognized a 28.5Kda protein which was defined as SEB by Western-blot analysis which indicated McAbs with high specificities were successfully established.Conclusion McAbs against SEB were successfully obtained and could be used to detect SEBin hospital and test department.
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