Antibody immobilisation on fibre optic TIRF sensors

Biosens Bioelectron. 2003 Nov 15;19(2):85-93. doi: 10.1016/s0956-5663(03)00173-8.

Abstract

A study of antibody immobilisation techniques on quartz and fibre optic surfaces for immunosensors has been carried out. Methods of covalent antibody immobilisation which have not previously been applied to optical fibres were investigated, and compared with classical methods found in the literature. Preliminary experiments on covalent immobilisation methods on planar quartz surfaces were conducted to enable us to choose the most suitable protein immobilisation technique for sensor applications. The immobilisation studies were directed in particular towards obtaining a high density of binding sites for the analyte of interest. Two of the most promising methods, antibody immobilisation on surfaces coated with dextran based hydrogel and F(ab')-SH fragments bound to silanised glass, which resulted in surface densities of active sites of above 0.45 pmol/cm2, were selected for further experiments on a fibre optic total internal reflection fluorescence immunosensor and gave satisfactory responses to changes in analyte concentrations of the order of 10(-8) M. The efficiency of polar organic solvents, such as dimethylsulfoxide, in dissociating the antigen-antibody complex and hence to regenerate the immunosensor surface was also evaluated.

Publication types

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

MeSH terms

  • Adsorption
  • Animals
  • Antibodies / analysis
  • Antibodies / chemistry*
  • Antibodies / immunology
  • Antigen-Antibody Complex / analysis
  • Antigen-Antibody Complex / immunology
  • Biosensing Techniques / instrumentation*
  • Biosensing Techniques / methods
  • Coated Materials, Biocompatible / chemical synthesis
  • Coated Materials, Biocompatible / chemistry
  • Electrodes*
  • Equipment Design
  • Equipment Failure Analysis
  • Fiber Optic Technology / instrumentation*
  • Humans
  • Immunoassay / instrumentation
  • Immunoassay / methods*
  • Immunoglobulin G / analysis*
  • Immunoglobulin G / immunology
  • Rabbits
  • Serum Albumin / analysis*
  • Serum Albumin / immunology
  • Spectrometry, Fluorescence / instrumentation*
  • Spectrometry, Fluorescence / methods

Substances

  • Antibodies
  • Antigen-Antibody Complex
  • Coated Materials, Biocompatible
  • Immunoglobulin G
  • Serum Albumin