[Study on screening mimicking epitope of ochratoxin A from phage display peptide library].
OBJECTIVE
To screen phage particles capable of mimicking ochratoxin A and establish immunoassay for ochratoxin A with it.


METHODS
An anti-ochratoxin A monoclonal antibody was used as ligand. Biopanning was done to screen the mimicking epitope from a phage random 7 peptide library. This library was displayed as a fusion protein with the coat protein III of filamentous phage M13. The positive clones were identified by ELISA, the inserted amino sequences were deduced by DNA sequencing.


RESULTS
After four rounds of panning, 11 positive clones can bind to the antibody, and the binding can be blocked the free ochratoxin A. The common amino sequence of the mimicking epitope is IRPMVXX. A competitive ELISA immunoassay was established with clone P2, the linear range of the inhibition curves is between 200 pg/ml and 8000 pg/ml, the detecting limitation is 150 pg/ml.


CONCLUSIONS
The phage display technique can be successfully applied to screen the mimicking epitope of ochratoxin A. The acquired phages may be used as the surrogate of the toxin to establish the immunoassay.
Liu
Ren-Rong
RR
Key Laboratory of Food Science of MOE, Nanchang University, Nanchang 330047, China.
Yu
Zhou
Z
He
Qing-Hua
QH
Wang
Xi
X
Xu
Yang
Y
D000911:Antibodies, Monoclonal / Q000276:immunology*; D001665:Binding Sites; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology*; D018716:Molecular Mimicry; D009793:Ochratoxins / Q000276:immunology*; D019151:Peptide Library
D000911:Antibodies, Monoclonal; D000939:Epitopes; D009793:Ochratoxins; D019151:Peptide Library; C025589:ochratoxin A