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[Mimic epitope of aflatoxin B1 screened by phage display technique]
Shengliang Deng1, Yang Xu, Renrong Liu
1Sino-German Joint Reserch Institute, the Key Laboratory of Food Science of MOE, Nanchang University, Nanchang 330047, China.
Objective:
To screen mimic epitope from phage-display random peptide libraries as sources of peptides that mimic the binding of aflatoxin B1 to monoclonal antibodies raised against the toxin and establish immunoassay for aflatoxin B1.
Methods:
The monoclonal antibody against the aflatoxin B1 was used as ligand to screen the binding peptide from the Ph.D.-7 peptide library with phage display and the specificity of colons were identified by ELISA. The peptide sequences of positive phage clones were determined and analyzed by DNA sequencing.
Results:
After four rounds of panning, the binding peptides were screened from random peptide libraries. Through indirect competitive ELISA, 4 positive clones could inhibited aflatoxin B1 and HXXDPXH was characterized by DNA sequencing, X is random amino acid. Then competitive ELISA was established for clone phage 10, the linear ranges of the inhibition curves were between 100 pg/ml and 2000 pg/ml, the detecting limit was 50 pg/ml.
Conclusion:
The mimic epitope of aflatoxin B1 antigen was obtained successfully by the phage display technigue and it is effective substitutes for aflatoxin B1-protein conjugate in the immunoassay.
