An immunoassay for ochratoxin A without the mycotoxin

Phage display peptide libraries allow the selection of new molecules capable of mimicking the structural and functional features of native proteins or chemicals. This technology can be applied to developing new reagents, which may serve as the surrogate of the original objects. In order to screen peptides capable of mimicking ochratoxin A (OTA) in the interaction with anti-OTA monoclonal antibody (McAb) and establish the immunoassay for OTA, an anti-OTA McAb was used as the target for panning-elution selection from a phage random seven-peptide library. After four rounds of panning, 11 phages were found to be able to mimic OTA in binding with the antibody. Enzyme-linked Immunosorbent Assay (ELISA) for detecting OTA was established with the phages. In the most sensitive assay, the linear range of the inhibition curve was 200–8000 pg/ml; the detection limit was 150 pg/ml. The inserted peptide sequences were deduced by DNA sequencing. The common amino acid residue sequence was IR(V)PMV(L)XX (X is any amino acid residues), which was verified by two synthesized peptides. The results demonstrated that those phage peptides could be used as the surrogate of OTA to establish the immunoassay.