鸡源性抗FB_1噬菌体单链抗体库的构建与鉴定

Construction and identification of a phage-displayed chicken sc Fv antibody library against fumonisin B_1

目的构建免疫鸡源性噬菌体单链抗体基因文库,筛选鉴定抗伏马菌素B1(fumonisin B1,FB1)的单链抗体。方法制备抗原免疫来亨母鸡,提取脾脏RNA反转录后通过PCR扩增获得单链抗体编码基因,构建单链抗体基因文库,噬菌体展示技术筛选抗FB1单链抗体,并利用表达ELISA检测抗体的亲和力。结果构建的单链抗体库的质量较高,通过噬菌体展示淘选和ELISA鉴定获得5个抗FB1的单链抗体。结论成功构建鸡源性抗FB1噬菌体单链抗体库,并鉴定获得抗FB1的单链抗体,表明鸡源性噬菌体单链抗体库可用于分离FB1等小分子半抗原的特异单链抗体。

To construct a phage-displayed single-chain variable fragment（sc Fv） antibody library and screen chicken-derived sc Fv antibodies against fumonisin B1（FB1）, Leghorn chickens were immunized with prepared antigen. Total RNA was extracted from spleen and sc Fv coding genes were reverse-transcripted and amplified.Then, FB1-specific antibodies were isolated by phage display technology from the library. The selected sc Fv genes were induced in Escherichia coli to express proteins, and the protein binding capacities were determined by enzyme-linked immunosorbent assay（ELISA）. The results showed that the constructed sc Fv antibody library had a good positive ratio, and five anti-FB1 sc Fv antibodies were obtained by phage-display panning and ELISA identification. In conclusion, a phage-displayed sc Fv antibody library is successfully constructed and chicken sc Fv antibodies against FB1 are selected, indicating that chicken-derived phage-displayed sc Fv antibody library could be utilized to isolate specific sc Fv antibodies against small molecules（haptens）.