牛乳铁蛋白单克隆抗体的制备及双抗夹心ELISA检测方法的建立  

Preparation of Anti-bovine Lactoferrin Monoclonal Antibodies and Development of Sandwich ELISA for the Detection of Bovine Lactoferrin

作　　者：项开合[1] 张乃生[2] 

机构地区：[1]辽东学院农学院,辽宁丹东118000 [2]吉林大学动物医学学院,吉林长春130062

出　　处：《中国兽医杂志》2014年第10期34-36,共3页Chinese Journal of Veterinary Medicine

摘　　要：本试验以牛乳铁蛋白为抗原免疫BALB/c小鼠,应用淋巴细胞杂交瘤技术制备单克隆抗体,获得了3株稳定分泌乳铁蛋白单克隆抗体的杂交瘤细胞株,分别命名为3C6、3F8和4D3,最后用相对亲和力最强的3C6所产生的抗体作为包被抗体,用辣根过氧化物酶标记过的多克隆抗体作为酶标抗体,最终建立了用于检测牛乳汁中乳铁蛋白含量的双抗夹心ELISA检测方法,该法对牛乳铁蛋白最低检测限为4 ng/m L,最适检测范围为4~256 ng/m L。This study was aimed to establish double antibody sandwich ELISA method for detection of bovine lactoferrin（LF）.Using bovine lactoferrin as antigen to immunize BALB/c mice and by lymphocyte hybridoma technique,monoclonal antibodies（Mc Ab）against bovine LF were obtained.Three strains of hybridoma cells were established and named 3C6,3F8 and 4D3.With purified mouse anti-bovine Mc Ab against LF which have the highest affinity and generated by 3C6 as a coated antibody,and rabbit HRPlabeled Pc Ab as a second detection antibody,double antibody sandwich ELISA was established for detection of bovine LF in milk.The lowest detectable limit of ELISA was 4ng/m L.The range of detection was 4~256 ng/m L.

关 键 词：乳铁蛋白 单克隆抗体 多克隆抗体 双抗夹心ELISA检测方法 

分 类 号：S854.43[农业科学—临床兽医学]