抗软海绵酸单克隆抗体的制备  

Preparation of monoclonal antibody against okadaic acid

作　　者：孙迎春[1] 陈杰[1] 刘军[2] 严建忠[2] 刘宁[3] 曹雅明[2] 潘国伟[3] 刘振林[1] 

机构地区：[1]中国医科大学公共卫生学院尘肺研究室,沈阳110001 [2]中国医科大学基础医学院免疫学教研室 [3]辽宁省疾病预防控制中心环境流行病科

出　　处：《中国公共卫生》2004年第2期186-188,共3页Chinese Journal of Public Health

基　　金：辽宁省科学技术基金资助项目 (992 2 80 0 1 )

摘　　要：目的　利用B淋巴细胞杂交瘤技术制备抗软海绵酸 (OA)的单克隆抗体 (McAb)。方法　用碳化二亚胺法将半抗原OA与牛血清白蛋白 (BSA)制备成完全抗原OA -BSA ,用OA -BSA免疫BALB/c小鼠 ,使小鼠脾细胞产生抗OA的抗体。在聚乙二醇的作用下 ,使脾细胞与小鼠骨髓瘤细胞融合在一起 ,融合后的细胞用HAT选择培养 ,最后用ELISA法筛选出分泌抗OAMcAb的杂交瘤细胞株。结果　用碳化二亚胺法成功地将微量半抗原OA与BSA和卵清白蛋白 (OVA)制成了完全抗原OA -BSA和OA -OVA。免疫小鼠血清中抗OA的抗体呈现阳性反应 ,表明小鼠脾细胞产生了抗OA的抗体。经 3次抗体检测筛选出 3株分泌抗OAMCAb的杂交瘤细胞株。结论　抗OAMcAb的成功制备为建立快速、经济的软海绵酸ELISA检测方法打下了坚实的物质基础 ,也为其它贝毒素检测方法的研究提供了可借鉴的经验。Objective To prepare monoclonal antibodies (McAb) against okadaic acid (OA) using B lymphocytes hybrid technique.Methods To prepare immunogen of OA-BSA,hapten of OA is coupled to bovine serum albumin (BSA) according to carbodiimide method.Splenic cells of mice immunized by OA-BSA produce antibodies against OA.The cell fusion is carried out by mixing splenic cells with the myeloma cells using polyethylene glycol (PEG).After the cell fusion,hybridoma cells are screened in a HAT medium.Hybridoma cells strains secreting monoclonal antibodies against OA are selected out by using enzyme-linked immunosorbent assay(ELISA).Results According to carbodiimide method,antigens of OA-BSA and OA-OVA are successfully prepared from minute amount hapten of OA.Antibodies against OA are positive in sera of mice,which shows splenic cells can secrete antibodies against OA.After three times detection of antibodies,three hybridoma cells strains secreting monoclonal antibodies against OA are obtained.Conclusion The successful preparation of monoclonal antibodies against OA supply a firm substantial foundation for developing an ELISA method that can determine OA as quickly and economically as possible in our country and furnish a experience that can be used as a reference on developing a method by which other shellfish toxins can be determined.

关 键 词：软海绵酸 单克隆抗体 ELISA 

分 类 号：R392.33[医药卫生—免疫学] Q547[医药卫生—基础医学]