title: Development of a single-chain variable fragment antibody-based enzyme-linked immunosorbent assay for determination of fumonisin B₁ in corn samples.
abstract: BACKGROUND
  Fumonisin B1 (FB1 ) 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 FB1 in corn samples.
  
  
  RESULTS
  Anti-FB1 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-FB1 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).
  
  
  CONCLUSIONS
  This scFv-based ELISA could be used as an efficient screening method for routine monitoring the residues FB1 in corn samples.
authors.affiliation: State Key Laboratory of Food Science and Technology, Sino-Germany Joint Research Institute, Nanchang University, Nanchang, 330047, China.
authors.forename: Long
authors.initials: L
authors.lastname: Zou
authors.forename: Yang
authors.initials: Y
authors.lastname: Xu
authors.forename: Yanping
authors.initials: Y
authors.lastname: Li
authors.forename: Qinghua
authors.initials: Q
authors.lastname: He
authors.forename: Bo
authors.initials: B
authors.lastname: Chen
authors.forename: Dan
authors.initials: D
authors.lastname: Wang
mesh_terms: D000911:Antibodies, Monoclonal / Q000235:genetics; D003001:Cloning, Molecular; D004032:Diet; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005506:Food Contamination / Q000032:analysis*; D037341:Fumonisins / Q000032:analysis*; D005670:Fusarium / Q000378:metabolism; D006801:Humans; D012639:Seeds / Q000382:microbiology; D057127:Single-Chain Antibodies / Q000235:genetics; D003313:Zea mays / Q000382:microbiology
chemical_list: D000911:Antibodies, Monoclonal; D037341:Fumonisins; D057127:Single-Chain Antibodies; C056933:fumonisin B1; C119496:tositumomab I-131
keywords: ELISA; corn; fumonisin B1; monoclonal antibody; phage display; single-chain variable fragment