A highly stable, sensitive, regenerable and rapid immunoassay for detecting aflatoxin B1 in corn incorporating covalent AFB1 immobilization and a recombinant Fab antibody

Talanta. 2013 Oct 15;115:329-35. doi: 10.1016/j.talanta.2013.05.012. Epub 2013 May 14.

Abstract

A highly robust immunoassay applicable for the detection of aflatoxin B1 (AFB1) using a Fab antibody fragment was developed. A key factor was the use of covalently immobilized AFB1 which allowed an almost three fold increase in sensitivity, reduced assay time and regeneration with retention of binding capacity. Various factors that might affect the sensitivity of the assay such as pH, organic solvents, storage stability and wash stringency were critically evaluated. It was also demonstrated that the assay was applicable for determination of AFB1 in corn samples at concentration within the European union regulatory limits.

Keywords: Amide bond; Covalent immobilization; Enzyme-linked immunosorbent assay (ELISA); Passive adsorption; Re-usability.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aflatoxin B1 / analysis*
  • Antibodies / chemistry*
  • Antibodies, Immobilized / biosynthesis
  • Antibodies, Immobilized / chemistry*
  • Antibody Affinity
  • Antibody Specificity
  • European Union
  • Hydrogen-Ion Concentration
  • Immunoassay*
  • Immunoglobulin Fab Fragments / biosynthesis
  • Immunoglobulin Fab Fragments / chemistry*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Solvents
  • Zea mays / chemistry*

Substances

  • Antibodies
  • Antibodies, Immobilized
  • Immunoglobulin Fab Fragments
  • Recombinant Proteins
  • Solvents
  • Aflatoxin B1