Construction and identification of phage ScFv library against deoxynivalenol.

The VH and VL gene fragments were first amplified from cDNA of hybridoma cell line specific to deoxynivalenol,integrated as ScFv with a pair of linker primers,cloned into phagemid vector pCANTAB-5E,and then transformed into Escherichia coli TG1.To produce phage-display ScFv,the helper phage M13KO7 was added to the cultures when bacteria were grown up to the log-phase,in which ScFv antibody was displayed in N-terminus of the bacteriophage coat protein pⅢ.The phage antibody contained 1.1×105 independent clones,and the titer of which was about 1.3×108 pfu.Enzymatic digestions of 10 random clones showed that the recombinant plasmids contained an intact ScFv gene.These results indicated that the phage antibody library was constructed successfully.