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

作　　者：龙军[1] 陈清[2] 俞守义[2] 

机构地区：[1]第一军医大学珠江医院检验科,广州510282 [2]广州第一军医大学热卫系流行病教研室

出　　处：《中国公共卫生》2005年第1期25-26,共2页Chinese Journal of Public Health

基　　金：军队指定性项目(01L050)

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。Objective To prepare Anti-staphylococcal EnterotoxinB(SEB) Moncolonal antibodiesMethods 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 IgGlsubclass except two strains(IgG2a)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 SEB in hospital and test department.

关 键 词：金黄葡萄球菌 肠毒素 单克隆抗体 腹水 

分 类 号：R155.31[医药卫生—营养与食品卫生学]