【摘要】 本研究使用噬菌体抗体库技术筛选针对庆大霉素(gentamicin)的单链抗体。以抗庆大霉素杂交瘤细胞株(2A3)为基因来源构建scFv基因,连接至pCANTAB5E噬菌粒载体,通过电击转化TG1(Escherichia coli TG1)构建了库容量为6.5×106噬菌体单链抗体库。抗庆大霉素噬菌体单链抗体库进行三轮富集筛选,通过Phage-ELISA技术成功筛选出了9个抗庆大霉素噬菌体阳性克隆,为开发新型残留检测用抗体奠定了基础。更多还原

【Abstract】 In this study,we constructed a phage display antibody library specific for gentamicin(GEN) to get the anti-getamicin single chain antibody(scFv).We used the anti-gentamicin hybridoma(2A3) as a source of gene to produce full-length scFv fragment.The full-length scFv fragment was ligated into the phagemid vector pCANTAB5E.Then the phagemid was transformed to E.coli TGl cells by electroporation.The size of antibody libraries was 6.5×106.The primary phage display library was riched by stabilized antigen immunoaffinity panning with GEN-OVA conjugates.9 clones congtained insert and showed GEN-positive in phage-ELISA,which lay the foundation for further immunological analysis.更多还原