Research on Panning Methods of Single-Domain Heavy-Chain Antibody Fragments for Aflatoxin B1 from Non-Immune Library

Abstract:

The objective of this study was tocompare the efficiency of two elute strategies in biopanning of single-domain heavy-chain antibody(HCAbs) phage display library against low molecular hapten,Aflatoxin B1(AFB1).For this,the artificial antigen AFB1-BSA and AFB1-OVA were used as ligand to screen the binding phages from an Alpaca non-immune library by antigen-coated solid phase selection.The binding phages were directly eluted with Gly-HCl or competitively eluted with AFB1 solution.The eluants were tittered separately.It was found that the recovery rate of Gly-HCl is 5 to 10 times higher than that of competition method.Twenty colons randomly picked from each eluant were characterized by phage-ELISA.Eight positive colons were isolated from the eluant by the competition method;however,no positive colons were isolated by Gly-HCl method.The sequencing data show that all the colons are encoding single-domain HCAbs.