鼠源抗AFB1噬菌体单链抗体库的构建与鉴定  

Construction and identification of mouse Anti-AFB1 phage single-chain antibody library

作　　者：裴世春[1] 孙大庆[2] 郭德军[1] 宋薇[1] 蒋琛[1] 

机构地区：[1]黑龙江八一农垦大学食品学院,黑龙江大庆163319 [2]黑龙江八一农垦大学黑龙江省农产品加工工程技术研究中心,大庆163319

出　　处：《齐齐哈尔大学学报（自然科学版）》2010年第6期76-80,共5页Journal of Qiqihar University(Natural Science Edition)

基　　金：黑龙江省教育厅项目(1151hz025)

摘　　要：研究以AFB1-BSA免疫的小鼠脾脏细胞为试验材料,利用RT-PCR技术,克隆了抗AFB1抗体重链和轻链可变区基因VH和VL,利用连接肽(Gly4Ser)3将VH和VL链接成单链抗体基因scFv,通过噬菌体展示载体pCANTAB-5E携带将其电转化E.coli TG1,经氨苄青霉素平板筛选,构建了库容为2.13×109 cfu/μg DNA的噬菌体单链抗体库,抗体库的克隆阳性率达到100%,且多样性良好,为高活性抗AFB1单链抗体的筛选提供了材料基础。The immunitied spleen cells of mouse by AFB1-BSA were used for test materials,resisted AFB1 antibody heavy chain VH and light chain gene VL were cloned by using RT-PCR,the linker peptide（Gly4Ser）3 was used to link VH and VL into a single chain gene scFv,through phage vectors carrying the pCANTAB-5E transformated into E.coli TG1 by electricity,adopted ampicillin flat screening,contenting 2.13×109 cfu/μg DNA of phage single-chain antibody libraries were constructed,the positive clone of antibody library reached 100%,provided the materials base for screening of highly active anti-AFB1 antibody.

关 键 词：黄曲霉毒素B1 单链抗体 RT-PCR 

分 类 号：R446.61[医药卫生—诊断学]