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Development of a single‐chain variable fragment antibody‐based enzyme‐linked immunosorbent assay for determination of fumonisin B 1 in corn samples
Author(s) -
Zou Long,
Xu Yang,
Li Yanping,
He Qinghua,
Chen Bo,
Wang Dan
Publication year - 2014
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/jsfa.6505
Subject(s) - fumonisin b1 , single chain variable fragment , fumonisin , immunoglobulin light chain , mycotoxin , chemistry , microbiology and biotechnology , antibody , phage display , monoclonal antibody , biology , food science , genetics
BACKGROUND Fumonisin B 1 ( FB 1 ) is a cancer‐promoting mycotoxin produced by Fusarium species and one of the major food‐borne toxins in corn and corn products. The objective of this study was to produce a single‐chain variable fragment ( scFv ) antibody for determination of FB 1 in corn samples . RESULTS Anti‐ FB 1 monoclonal antibodies were obtained via the hybridoma technique. Specific heavy‐ and light‐chain variable fragments were amplified with degenerate primers and constructed into scFv antibody fragments by splice overlap extension polymerase chain reaction with linker sequences. The resulting scFv DNA fragments were cloned into the phagemid pHEN1for selection and identification of functional scFv fragment by phage display. Prokaryotic expression vector pET22b‐scFv was constructed to prepare anti‐ FB 1 scFv antibody for establishment of indirect competitive ELISA . The detection capability ( CCβ ) of the scFv ‐based ELISA was 15.00 µg kg −1 , and the limit of detection was 8.32 µg kg −1 . The recoveries and coefficients of variation were 86.74–107.34% and 9.72–14.03%, respectively. In addition, the determined results of 30 naturally contaminated corn samples by the scFv ‐based ELISA are in agreement with the findings of high‐performance liquid chromatography ( R 2 = 0.97) . CONCLUSION This scFv‐based ELISA could be used as an efficient screening method for routine monitoring the residues FB 1 in corn samples. © 2013 Society of Chemical Industry