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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。
Objective To construct a library of single-chain antibody against chicken-derived phage avian influenza virus and screened the single chain antibody against fumonisin B1 (FB1). Methods The antigens were immunized to Henheng chicken, RNA of the spleen was reverse transcribed, and the single-chain antibody gene was amplified by PCR. Single-chain antibody gene library was constructed. The anti-FB1 single-chain antibody was screened by phage display technique. Affinity. Results The constructed single-chain antibody library was of high quality. Five anti-FB1 scFv antibodies were obtained by phage display panning and ELISA. Conclusion The chicken-derived anti-FB1 phage scFv library was successfully constructed and the anti-FB1 scFv was identified. The results showed that the chicken-derived phage scFv library can be used to isolate specific single-chain antibodies against small molecule haptens such as FB1.