【摘要】 为制备捻转血矛线虫(Haemonchus contortus)天然H11蛋白的单克隆抗体，以伴刀豆球蛋白(Concanavalin A,Con A)凝集素提取的天然H11蛋白为免疫原，免疫4～6周龄雌性BALB/c小鼠，3次免疫后分离小鼠脾细胞并将其与SP2/0细胞进行融合，通过间接ELISA方法筛选，获得2株能稳定分泌抗H11蛋白单克隆抗体的杂交瘤细胞株，分别命名为A1E3和A10E1。通过亚型鉴定和免疫学分析显示，2株单克隆抗体的重链均属于IgG1,轻链均为κ型，并且这2株单抗均能识别虫体天然抗原H11。免疫组化定位和体外幼虫发育抑制试验表明，mAb A1E3可以定位到成虫的肠道微绒毛部位，该抗体能够抑制四期幼虫的生长发育。单抗的成功制备不仅为研究H11蛋白保护性抗原表位以及开发表位疫苗奠定基础，也为单抗治疗动物捻转血矛线虫病提供了潜在的应用前景。更多还原

【Abstract】 To construct monoclonal antibodies against Haemonchus contortus native H11 protein.In this study, five 4-6 weeks female BALB/c mice were immunized with native H11 protein extracted from adult worms by Concanavalin A lectin.Spleen cells were isolated and fused with SP2/0 cells after 3 times of immunization.Two hybridoma cell lines, named A1E3 and A10E1,which could stably secrete monoclonal antibodies against H11 protein were obtained.The subtype identification and immunological analysis showed that the heavy chain of the two monoclonal antibodies belonged to IgG1 and the light chain was κ type, and both monoclonal antibodies recognized the natural antigen H11.Immunohistochemical localization and larval developmental inhibition test in vitro showed that the mAb A1E3 could be localized to the intestinal microvilli of the adult worm, and that the antibody can inhibit the growth of the fourth-stage larvae.The successful production of two monoclonal antibodies not only lays the foundation for the study of protective antigenic epitopes of the H11 protein and the development of epitope vaccines, but also provides a potential application of the monoclonal antibody for the treatment of haemonchusis in animals.更多还原