为制备抗大肠杆菌不耐热肠毒素B亚单位(LTB)的单克隆抗体(MAb),本研究以纯化的原核重组LTB蛋白(rLTB)免疫6周龄BALB/c小鼠,细胞融合后经间接ELISA筛选,获得两株能够稳定分泌抗LTB抗体的杂交瘤细胞株2F11和4H2.抗体亚类鉴定均为IgG1类.Western blot结果显示,这两株MAb均可以与五聚体形式存在的rLTB蛋白发生特异性结合,表明两株MAb可以用于天然LTB的检测.本实验为LTB的检测与应用提供了实验依据.%To prepare the monoclonal antibody (MAb) against E.coli heat-labile enterotoxin B submit (LTB), 6 weeks BALB/c mice were immunized 3 times with purified recombinant LTB protein (rLTB) expressed in E.coli. Two hybridoma clones secreting antibody against rLTB were obtained by fusing the splenocytes from immunized mice with SP2/0 myeloma cells. With purified rLTB as detecting antigen, MAbs against rLTB were prepared and positive hybridoma clones were screened by tndirect-ELISA. The 2 MAbs belonged to IgG1. The result of western blot showed 2 MAbs were able to specifically recognize pentamer LTB protein, and confirmed the MAbs had practical application.
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