牛乳铁蛋白LF单克隆抗体的制备

Preparation of Monoclonal Antibodies Against LF of Bovine

制备牛乳铁蛋白单克隆抗体,研究利用标准LF蛋白作为免疫原,免疫六周龄Balb/c雌性小鼠,运用淋巴细胞杂交瘤技术,通过间接ELISA筛选,采用有限稀释法,经过3次克隆筛选,获得两株稳定分泌抗LF蛋白的杂交瘤细胞株,命名为:L1,并进行了亚类鉴定。结果表明:该杂交瘤细胞培养上清抗体效价L1为1:4.56×104;腹水L1为1:5.21×106;L1分泌的单克隆抗体为IgG1型,轻链为λ链抗体。

Six-week-old female Balb/c mice were immunized with LF protein.Two hybridoma cell lines named L1 and L2 respectively,were obtained by using hybridoma technique,limiting dilution and three-time subclone.All the cell lines was identifined and analyzed by indirect ELISA,the specific reaction of secreted monoclonal antibody（McAb） against LF is stable.The isotypes of McAbs secreted by L1,L1 were IgG1,belonged to λ chain.The antibody titers of L1 hybridoma cells culture supernate was 1：4.56×104;abdominal dropsy of L1 was 1：5.21×106,anabiosis the hybridoma cell,to detect supernatant and abdominal dropsy of hybridoma cells by ELISA,compared the OD fig,the result indicated that four generations titer of supernatant and abdominal dropsy was descend slightly.