Objective To prepare nanobodies of aflatoxin B1(AFB1)by using fully synthesized nanobody library.Methods The complete antigen of AFB1 was prepared and screened in 4 rounds relying on the fully synthesized nanobody library,and the captured high-affinity antibody gene was transformed into the prokaryotic expression system for expres-sion to analyze the specificity and affinity.In addition,the three-dimensional structure of the antibody G5-AFB1 was con-structed to analyze the binding characteristics of the two.Results On the basis of AFB1 complete antigen preparation,specific phage was obviously enriched with the increase of rounds,and finally a positive monoclonal G5 was obtained,which was identified as having effective concentration 50％(EC50)of 0.953 μmol/L and an half maximal inhibitory con-centration(IC50)of 43 μmol/L.In addition,the G5-AFB1 complexes not only formed a stable hydrogen bond,but also possessed better hydrophobicity and electrostatic effects.Conclusion The ability to screen good AFB1 antibodies by fully synthetic nanobody library provides a good basis for AFB1 detection and treatment,which confirms that phage display technology as well as fully synthesized nanobody library have a large potential for development in the preparation of small molecule biotoxin antibodies.
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