不同杂交瘤细胞株单链抗体的构建及性质比较  

Constrcution and Comparison of Anti-AFB1 ScFv from Different Hybridoma Cell Lines

作　　者：胡莉 刘爱平[2] 王小红[3] HU Li;LIU Aiping;WANG Xiaohong(College of Biological and Food Engineering,HuangHuai University,Zhumadian 463000,China;College of Food Science,Sichuan Agricultural University,Yaan 625014,China;College of Food Science and Technology,Huazhong Agricultural University,Wuhan 430070,China)

机构地区：[1]黄淮学院生物与食品工程学院,河南驻马店463000 [2]四川农业大学食品学院,四川雅安625014 [3]华中农业大学食品科技学院,湖北武汉430070

出　　处：《食品与生物技术学报》2019年第11期70-77,共8页Journal of Food Science and Biotechnology

基　　金：国家自然科学基金项目（31271876）

摘　　要：不同的杂交瘤细胞株,分泌的抗黄曲霉毒素单克隆抗体的灵敏度也大不相同,通过筛选获得三株杂交瘤细胞,分别命名为2C10、2E6和2H1,其中2C10分泌的单克隆抗体在酶联免疫吸附(ELISA)试验中具有最高的灵敏度,2E6次之,2H1再次之。为了能够更好的展现和理解抗体和抗原之间的反应关系,以及最大程度的提高重组抗体的灵敏度,实验通过构建3株细胞株的单链抗体,并在E.coli BL21(DE3)中表达纯化单链抗体蛋白质,采用竞争ELISA法展现了不同的单克隆抗体与所对应的单链抗体灵敏度之间的关系。结果表明,灵敏度最好的单克隆抗体2C10所对应的单链抗体,其灵敏度也是最好的,2E6的次之,2H1的再次之。同时实验也表明,人工合成的单链抗体的灵敏度较其所对应的单克隆抗体的灵敏度要下降许多。Anti-AFB1 Monoclonal antibody secreted by the differeent hybridoma cell lines have different sensitivity,three different hybridoma cell lines named 2C10,2E6 and 2H1 were screened and studied,the result of enzyme-linked immunosorbent(ELISA)test showed that the sequence of the sensitivity of monoclonal antibodies secreted by 2C10,2E6 and 2H1 was 2C10>2E6>2H1.Three anti-AFB1 scFvs from different hybridoma cell lines were constructed and expressed in E.coli BL21(DE3)respectively in order to better show and understanding the relationship between antigen and antibody and improve the sensitivity of recombinant antibodies at the greatest extent.The ELISA method was used to detect the sensitivity relationship between the different monoclonal antibodies and anti-AFB1 scFvs after protein expression and purification.The result showed that the sequence of the sensitivity of anti-AFB1 scFvs was scFv-2C10>scFv-2E6>scFv-2H1,and showed that the sensitivity of monoclonal antibodies were much better than the corresponding anti-AFB1 scFvs.

关 键 词：黄曲霉毒素B1 单克隆抗体 单链抗体 ELISA 

分 类 号：Q786[生物学—分子生物学]