Development of a single‐chain variable fragment antibody‐based enzyme‐linked immunosorbent assay for determination of fumonisin B1 in corn samples. (29th December 2013)

• Record Type: Journal Article
• Authors: Zou, Long; Xu, Yang; Li, Yanping; He, Qinghua; Chen, Bo; Wang, Dan
• Is Part Of: Journal of the science of food and agriculture. Volume 94:Number 9(2014:Jul. 15)
• Page Start: 1865
• Page End: 1871
• Publication Date: 2013-12-29
• Subjects: Food -- Periodicals; Agriculture -- Periodicals; 664
• DOI: 10.1002/jsfa.6505
• Languages: English
• ISSNs: 0022-5142

Abstract

<abstract abstract-type="main" id="jsfa6505-abs-0001"> <title>Abstract</title> <sec id="jsfa6505-sec-0001" sec-type="section"> <title>BACKGROUND</title> <p id="jsfa6505-para-0001"> <bold>Fumonisin B<sub>1</sub> (FB<sub>1</sub>) is a cancer‐promoting mycotoxin produced by <italic>Fusarium</italic> 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<sub>1</sub> in corn samples</bold>.</p> </sec> <sec id="jsfa6505-sec-0002" sec-type="section"> <title>RESULTS</title> <p id="jsfa6505-para-0002"> <bold>Anti‐FB<sub>1</sub> 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<sub>1</sub> scFv antibody for establishment of indirect competitive ELISA. The detection capability (CCβ) of the scFv‐based ELISA was 15.00 µg kg<sup>−1</sup>, and the limit of detection was 8.32 µg kg<sup>−1</sup>. 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 (<italic>R</italic><sup>2</sup> = 0.97)</bold>.</p> </sec> <sec id="jsfa6505-sec-0003" sec-type="section"> <title>CONCLUSION</title> <p id="jsfa6505-para-0003"> <bold>This scFv‐based ELISA could be used as an efficient screening method for routine monitoring the residues FB<sub>1</sub> in corn samples. © 2013 Society of Chemical Industry</bold> </p> </sec> </abstract>