抗庆大霉素噬菌体抗体库的构建和筛选

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6.5×106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。

Construction of Anti-gentamicin Phage Display scFv Libraries, Selection and Identification

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 6.5×106. 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.