巨型艾美耳球虫AMA1蛋白单克隆抗体的制备及其功能鉴定  

Preparation and Application of Monoclonal Antibodies against Apical Membrane Antigen 1 of Eimeria maxima

作　　者：罗伏兵 刘晓冬 梁琳[2] 刘贤勇 徐发荣 沈光年 马志军 索勋[3] 汤新明 LUO Fu-bing;LIU Xiao-dong;LIANG Lin;LIU Xian-yong;XU Fa-rong;SHEN Guang-nian;MA Zhi-jun;SUO Xun;TANG Xin-ming(Beijing Center for Animal Disease Control and Prevention,Beijing 102629,China;Institute of Animal Sciences,Chinese Academy of Agricultural Sciences,Beijing 100193,China;College of Veterinary Medicine,China Agricultural University,Beijing 100193,China)

机构地区：[1]北京市动物疫病预防控制中心,北京大兴102629 [2]中国农业科学院北京畜牧兽医研究所,北京海淀100193 [3]中国农业大学动物医学院,北京海淀100193

出　　处：《中国兽医杂志》2020年第11期34-36,I0007,共4页Chinese Journal of Veterinary Medicine

基　　金：北京市家禽产业创新团队(BAIC04-2020)。

摘　　要：为了研究巨型艾美耳球虫顶膜抗原1(AMA1)的细胞定位及功能,制备了抗AMA1的单克隆抗体并进行了鉴定。首先,用大肠杆菌(DE3)表达了巨型艾美耳球虫AMA1蛋白,蛋白经纯化后免疫小鼠,测定小鼠血清中AMA1抗体滴度。然后进行杂交瘤细胞的制备和克隆筛选,获得了16株分泌产生AMA1抗体的杂交瘤细胞株。最后用免疫印迹和间接免疫荧光试验对上述单克隆抗体进行鉴定。结果显示,编号为21和34的杂交瘤细胞株产生的抗体可识别巨型艾美耳球虫可溶性抗原中的AMA1组分;编号为34和35的抗体与巨型艾美耳球虫子孢子反应后,荧光信号集中于子孢子的顶质体和细胞膜部位。本试验制备的抗AMA1的单克隆抗体,为巨型艾美耳球虫生物学和免疫学研究提供了科学依据。To study the cellular localization and function of apical membrane antigen 1(AMA1)of Eimeria maxima,monoclonal antibodies against AMA1 were prepared and identified.The mice were immunized with recombinant AMA1 protein expressed by E.coli(DE3).Then,hybridoma cell preparation and clonal screening were performed,and 16 hybridoma cell lines producing AMA1 antibodies were obtained.Finally,the monoclonal antibodies were identified by Western Blot and indirect immunofluorescence assay.The results showed that the antibodies produced by the hybridoma cell lines numbered 21 and 34 could recognize the native AMA1 of the E.maxima soluble antigen.The antibodies numbered 34 and 35 reacted with E.maxima sporozoites with the fluorescence signal concentrated in the apical and cell membrane of the sporozoite.The monoclonal antibodies against AMA1 prepared in this study laid the foundation for the biological and immunological research of E.maxima.

关 键 词：巨型艾美耳球虫 顶膜抗原1 单克隆抗体 免疫印迹 免疫荧光 

分 类 号：R382.32[医药卫生—医学寄生虫学]