芝麻过敏原油质蛋白O leosins单克隆抗体的制备与鉴定  

Preparation and Identification of Monoclonal Antibodies against O leosins

作　　者：王媛媛[1] 邹菊[1] 刘志刚[1] 

机构地区：[1]深圳大学医学院过敏反应与免疫学研究所,广东深圳518060

出　　处：《食品科学》2013年第5期177-180,共4页Food Science

基　　金：深圳市产业发展专项资金创新平台建设计划重点实验室项目(SW201110010);深圳市科技计划深港创新圈项目(CX Q2008026);深圳大学科技创新团队项目(T200904);广东省大学生创新实验项目(2012026);深圳大学实验室开放基金项目(SD2012009)

摘　　要：利用细胞融合技术制备小鼠抗油质蛋白杂交瘤细胞,通过间接ELISA方法筛选稳定分泌特异性抗体的杂交瘤细胞株。获得10株杂交瘤细胞,分别命名为1B2、2D8、2F3、3A4、3D7、3F8、4A10、4F12、6A12、6E7,利用Ig亚类鉴定试剂盒鉴定各单克隆抗体的Ig亚型,除1B2、3D7、3F8为IgG1类外其余均为IgG2a类型。将杂交瘤细胞株注射入小鼠腹腔获取腹水,应用Protein A亲和层析法对腹水进行纯化。单抗效价有9株在2.0×105以上。ELISA和Western blotting分析表明这些单抗均能特异性识别芝麻过敏原油质蛋白。To prepare monoclonal antibodies against allergen O leosins from sesame seeds,BALB/c mice were immunized and the splenocytes of the immunized mice were fused with NS-1 myeloma cells by hybridoma technique.Ten hybridoma cell lines secreting McAbs against O leosins were obtained and classified as 1B2,2D8,2F3,3A4,3D7,3F8,4A10,4F12,6A12,6E7,1B2,3D7 and 3F8 were subtybes of IgG1 while 2D8,2F3,3A4,4A10,4F12,6A12,6E7 belonged to the subtype of IgG2a.The antibody titers were more than 2.0 × 105.The McAbs were then purified using affinity chromatography on immobilized Protein A and identified by their specificities,subtypes and titers with ELISA and Western blotting.The ten McAbs all recognized recombinant O leosins of sesame seeds.These monoclonal antibodies against O leosins were prepared successfully,which will facilitate to establish the detection of allergens in sesame seeds.

关 键 词：芝麻油质蛋白 单克隆抗体 特性鉴定 

分 类 号：TS201.6[轻工技术与工程—食品科学] R392.8[轻工技术与工程—食品科学与工程]