[Study on screening mimicking epitope of ochratoxin A from phage display peptide library]. 2005

Ren-Rong Liu, and Zhou Yu, and Qing-Hua He, and Xi Wang, and Yang Xu
Key Laboratory of Food Science of MOE, Nanchang University, Nanchang 330047, China.

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.

UI MeSH Term Description Entries
D009793 Ochratoxins Isocoumarins found in ASPERGILLUS OCHRACEUS and other FUNGI. Ochratoxin contaminated FOOD has been responsible for cases of FOODBORNE DISEASES. Ochratoxin
D004797 Enzyme-Linked Immunosorbent Assay An immunoassay utilizing an antibody labeled with an enzyme marker such as horseradish peroxidase. While either the enzyme or the antibody is bound to an immunosorbent substrate, they both retain their biologic activity; the change in enzyme activity as a result of the enzyme-antibody-antigen reaction is proportional to the concentration of the antigen and can be measured spectrophotometrically or with the naked eye. Many variations of the method have been developed. ELISA,Assay, Enzyme-Linked Immunosorbent,Assays, Enzyme-Linked Immunosorbent,Enzyme Linked Immunosorbent Assay,Enzyme-Linked Immunosorbent Assays,Immunosorbent Assay, Enzyme-Linked,Immunosorbent Assays, Enzyme-Linked
D000911 Antibodies, Monoclonal Antibodies produced by a single clone of cells. Monoclonal Antibodies,Monoclonal Antibody,Antibody, Monoclonal
D000939 Epitopes Sites on an antigen that interact with specific antibodies. Antigenic Determinant,Antigenic Determinants,Antigenic Specificity,Epitope,Determinant, Antigenic,Determinants, Antigenic,Specificity, Antigenic
D001665 Binding Sites The parts of a macromolecule that directly participate in its specific combination with another molecule. Combining Site,Binding Site,Combining Sites,Site, Binding,Site, Combining,Sites, Binding,Sites, Combining
D018716 Molecular Mimicry The structure of one molecule that imitates or simulates the structure of a different molecule. Antigenic Mimicry,DNA Mimicry,Mimicry, Molecular,Antigen Mimicry,Antigen Mimicries,Antigenic Mimicries,DNA Mimicries,Mimicries, Antigen,Mimicries, Antigenic,Mimicries, DNA,Mimicries, Molecular,Mimicry, Antigen,Mimicry, Antigenic,Mimicry, DNA,Molecular Mimicries
D019151 Peptide Library A collection of cloned peptides, or chemically synthesized peptides, frequently consisting of all possible combinations of amino acids making up an n-amino acid peptide. Phage Display Peptide Library,Random Peptide Library,Peptide Phage Display Library,Phage Display Library, Peptide,Synthetic Peptide Combinatorial Library,Synthetic Peptide Library,Libraries, Peptide,Libraries, Random Peptide,Libraries, Synthetic Peptide,Library, Peptide,Library, Random Peptide,Library, Synthetic Peptide,Peptide Libraries,Peptide Libraries, Random,Peptide Libraries, Synthetic,Peptide Library, Random,Peptide Library, Synthetic,Random Peptide Libraries,Synthetic Peptide Libraries

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