高亲和力抗藻毒素单链抗体的快速筛选  

Rapid Selection of scFv against Microcystins-LR

作　　者：党辉[1,2] 陈佩[3] 张秋香[1] 刘小鸣[1] 赵建新[1] 陈永泉[1] 张灏[1] 陈卫[1] 

机构地区：[1]江南大学食品学院,江苏无锡214122 [2]陕西师范大学食品工程与营养科学学院,西安710062 [3]陕西广播电视大学,西安710062

出　　处：《中国食品学报》2017年第10期274-279,共6页Journal of Chinese Institute Of Food Science and Technology

基　　金：江苏省产学研联合创新资金项目(BY2013015-25);农业科技创新与攻关项目:陕西省科学技术研究发展计划项目(2017NY-182)

摘　　要：蓝藻水华引发的微囊藻毒素(MC-LR)与肿瘤促进作用有直接关系,检测方法主要是色谱学方法和免疫学方法。以碳化二亚胺法将MC-LR与磁珠直接偶联,采用磁珠液相筛选法从噬菌体单链抗体库中筛选抗MCLR的单链抗体(scFv),结果成功富集、筛选到抗MC-LR的阳性克隆,阳性率达到95%。通过基因测序获得3株阳性克隆,其中与MC-LR结合活性最好的阳性克隆的IC_(50)为0.6 ng/m L。结果表明,将半抗原与磁珠直接偶联,可以灵敏、快速地从噬菌体抗体库中分离目标抗体。Microcystins-LR comes from blue-green alga bloom has a direct relationship with tumor promotion.The main detection methods of MC-LR are immunology and chromatography.In this study,MC-LR was coupled onto magnetic beads with a carbodiimide-mediated reaction.The beads-MC-LR was used to selects sc Fv against MC-LR from single-chain antibody phage display.The positive clones against MC-LR was successfully enriched and screened,and the positive rate achieved 95%.Three positive clones of different strains were obtained by gene sequenced,and IC50 of the positive clones for the best activity combined with MC-LR was 0.6 ng/m L.The results showed that the magnetic beads coupled with a hapten can be used to sensitively and rapidly select the target antibody from a phage antibody library.

关 键 词：磁珠 噬菌体单链抗体库 微囊藻毒素 筛选 单链抗体 

分 类 号：R392[医药卫生—免疫学]