Production and characterization of monoclonal antibody and its recombinant single chain variable fragment specific for a food-born mycotoxin, fumonisin B1

Bioprocess Biosyst Eng. 2010 Jan;33(1):109-15. doi: 10.1007/s00449-009-0350-9. Epub 2009 Jul 12.

Abstract

Fumonisin B(1) (FMB(1)) is a food-born mycotoxin produced by Fusarium moniliforme. Monoclonal antibody against FMB(1) (anti-FMB(1) mAb) was produced in the hybridoma DV9, which was established from a BALB/c mouse immunized with bovine serum albumin conjugated FMB(1) (FMB(1)-BSA). A competitive direct enzyme-linked immunosorbent assay (ELISA) showed that anti-FMB(1) mAb has about 10 ppb of minimum FMB(1) detection concentration and 220 ppb of 50% inhibition concentration (IC(50)). Much lower cross-reactivity of anti-FMB(1) mAb on ochratoxin A, aflatoxin B(1) and deoxynivalenol provided that anti-FMB(1) mAb was specific for FMB(1). The gene coding single chain variable fragment against FMB(1) (anti-FMB(1) scFv) was cloned from the hybridoma DV9 and was expressed in recombinant Escherichia coli. Insoluble anti-FMB(1) scFv required optimization of its refolding condition, and hence functional scFv was obtained. By using indirect ELISA, about 12-fold lower binding activity of anti-FMB(1) scFv on FMB(1)-BSA was obtained in comparison with that of the parental mAb.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aflatoxin B1 / chemistry
  • Aflatoxin B1 / immunology
  • Animals
  • Antibodies, Monoclonal / biosynthesis
  • Antibodies, Monoclonal / chemistry
  • Antibodies, Monoclonal / genetics
  • Antibodies, Monoclonal / immunology*
  • Antibody Affinity / genetics
  • Antibody Affinity / immunology
  • Antibody Specificity / genetics
  • Antibody Specificity / immunology*
  • Cattle
  • Cross Reactions / immunology
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Female
  • Fumonisins / chemistry
  • Fumonisins / immunology*
  • Fusarium / chemistry
  • Gene Expression
  • Mice
  • Mice, Inbred BALB C
  • Ochratoxins / chemistry
  • Ochratoxins / immunology
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Serum Albumin, Bovine / chemistry
  • Serum Albumin, Bovine / immunology
  • Single-Chain Antibodies / biosynthesis
  • Single-Chain Antibodies / chemistry
  • Single-Chain Antibodies / genetics
  • Single-Chain Antibodies / immunology*
  • Trichothecenes / chemistry
  • Trichothecenes / immunology

Substances

  • Antibodies, Monoclonal
  • Fumonisins
  • Ochratoxins
  • Recombinant Proteins
  • Single-Chain Antibodies
  • Trichothecenes
  • ochratoxin A
  • Serum Albumin, Bovine
  • fumonisin B1
  • Aflatoxin B1
  • deoxynivalenol