Photochemical attachment of biomolecules onto fibre-optics for construction of a chemiluminescent immunosensor

Boaz Leshem, Gadi Sarfati, Andres Novoa, Igor Breslav, Robert S. Marks

Research output: Contribution to journalArticlepeer-review

45 Scopus citations

Abstract

We report herein a simple and effective way to photochemically immobilize biomolecules onto a fibre-optic silica surface. The system is based on a photoreactive benzophenone derivative that is bound to SiO2 surfaces of the optical fibre via a silane anchor. The benzophenone derivative was 4-allyloxybenzophenone, synthesized by standard procedures that were later used to synthesize the 4-(3′-chlorodimethylsilyl) propyloxybenzophenone and 4-(3′-dichloromethylsilyl) propyloxybenzophenone by regular hydrosilation procedures. After silanization with the benzophenone derivatives, the fibres were immersed in a cholera toxin B subunit solution and illuminated with UV light (wavelength > 345 nm). As a result of the photochemical reaction, a thin layer of the antigen was covalently bound to the benzophenone-modified surface. The photochemically modified fibre-optics were then tested as immunosensors in the detection of cholera anti-toxin antibody and revealed through chemiluminescence measurements. A secondary antibody labelled with horseradish peroxidase acted as the marker for the cholera toxin antibody. A photo-electronic set-up was designed specifically to monitor the signal. The immunosensor system was shown to be both specific and sensitive. The lowest rabbit serum titre detected was 1:1 700 000.

Original languageEnglish
Pages (from-to)69-77
Number of pages9
JournalLuminescence
Volume19
Issue number2
DOIs
StatePublished - 1 Jan 2004

Keywords

  • Chemiluminescence
  • Cholera diagnostics
  • Fibre-optic immunosensor
  • Protein immobilization
  • Silane-benzophenone

ASJC Scopus subject areas

  • Biophysics
  • Chemistry (miscellaneous)

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