金黄色葡萄球菌肠毒素C3单克隆抗体的制备与鉴定  

Preparation and identification of monoclonal antibodies to staphylococcal enterotoxin C3

作　　者：王敏[1] 马瑞玉 李先平[1] 郑荣[1] 张婷婷[1] 曹虹[1] 

机构地区：[1]中南大学湘雅二医院检验科,湖南长沙410011 [2]中南大学湘雅医学院医学检验系,2007级湖南长沙410013

出　　处：《中国微生态学杂志》2014年第2期134-138,共5页Chinese Journal of Microecology

基　　金：湖南省科技计划项目(2013SK3048);湖南省卫生厅科研基金(132012024)

摘　　要：目的制备稳定分泌抗金黄色葡萄球菌肠毒素C3(SEC3)单克隆抗体的杂交瘤细胞株,并对单克隆抗体的性质进行鉴定。方法以SEC3重组蛋白免疫Balb/c小鼠,应用细胞融合技术将小鼠的脾细胞与SP2/0骨髓瘤细胞进行融合,经间接ELISA法检测筛选及2次有限稀释法克隆化培养,获得目的杂交瘤细胞株,并对其所产生的单克隆抗体进行效价、亲和常数及抗原识别表位等相关性质的鉴定。结果最终获得了两株能分泌单克隆抗体的杂交瘤细胞1C12和2A2,两者细胞培养上清的效价分别为1∶3 200和1∶1 600。经分析可知1C12细胞株的亲和力高于2A2细胞株,同时相加实验表明两个单克隆抗体识别抗原表位相同。结论单克隆抗体制备成功,为进一步完善肠毒素SEC3的临床检测奠定了基础。Objective To prepare and identify monoclonal antibodies against staphylococcal enterotoxin C3 （ SEC3 ）. Methods Hybridomas were developed by fusion of splenocytes from Balb/c mice immunized with puri- fied SEC3 protein and the myeloma cells （SPJ0）. They were screened by indirect enzyme-linked immunosorbent assay （ELISA） and isolated by limiting dilution twice. The characteristiess of monoclonal antibodies, such as titer, relative appetency, antigenic epitopes etc. were identified by ELISA. Results Two hybridomas producing mono- clonal antibodies against SEC3 were obtained finally and named 1C12 and 2A2 respectively. Their titers in culture supernatant were 1 ： 3200 and 1 ： 1600 respectively. The relative appetency of the monoclonal antibodies from 1 C12 was higher, and the monoclonal antibodies from the two hybridomas shared the same binding site of the anti- gen. Conclusion The successful preparation of monoclonal antibodies against SEC3 provides a basis for the fur- ther improvement of the detection of SEC3 in clinical practices.

关 键 词：金黄色葡萄球菌 SEC3 单克隆抗体 间接ELISA 

分 类 号：R378.11[医药卫生—病原生物学]