The impact of antibody/epitope affinity strength on the sensitivity of electrochemical immunosensors for detecting small molecules

Anal Bioanal Chem. 2013 Apr;405(11):3889-98. doi: 10.1007/s00216-013-6782-8. Epub 2013 Feb 27.

Abstract

A displacement immunoassay involves having a labelled analogue of the analyte (the epitope) already bound to the antibody. The presence of the analyte causes a competition for antibodies, and some of the antibodies dissociates from the epitope so that it can bind with the analyte. Herein, the influence of the affinity of the surface-bound epitope for the antibody on the sensitivity and selectivity of a displacement immunosensor is explored both theoretically and experimentally. An electrochemical immunosensor described previously, where the dissociation of antibodies from an electrode surface causes an increase in current from surface-bound ferrocene species, is used for this purpose. As expected, the ease and effectiveness of the bound antibody being displaced is inversely related to the affinity of the antibody to the surface-bound epitope relative to the analyte in solution as expected. However, if the affinity constant is too low, selectivity and/or sensitivity are compromised. Experimental results are qualitatively compared with a simple mass-action model.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / immunology
  • Antibody Affinity*
  • Antineoplastic Agents / analysis*
  • Antineoplastic Agents / immunology
  • Biosensing Techniques / methods
  • Electrochemical Techniques / methods
  • Enrofloxacin
  • Epitopes / chemistry
  • Epitopes / immunology*
  • Fluoroquinolones / analysis*
  • Fluoroquinolones / immunology
  • Immunoassay / methods*
  • Limit of Detection
  • Models, Molecular

Substances

  • Antibodies, Monoclonal
  • Antineoplastic Agents
  • Epitopes
  • Fluoroquinolones
  • Enrofloxacin