Preparation of Aflatoxin B1 Antibody Based on Fully Synthesized Nanobody Library
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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