We describe herein a newly developed optical microbiosensor for the diagnosis of hepatitis C virus (HCV) by using a novel photoimmobilization methodology based on a photoactivable electrogenerated polymer film deposited upon surface-conductive fiber optics, which are then used to link a biological receptor to the fiber tip through light mediation. This fiber-optic electroconductive surface modification is done by the deposition of a thin layer of indium tin oxide on the silica surface of the fiber optics. Monomers are then electropolymerized onto the conductive metal oxide surface; thereafter, the fibers are immersed in a solution containing HCV-E2 envelope protein antigen and illuminated with UV light (wavelength approximately 345 nm). As a result of the photochemical reaction, a thin layer of the antigen becomes covalently bound to the benzophenone-modified surface. The photochemically modified fiber optics were tested as immunosensors for the detection of anti-E2 protein antibody analyte that was measured through chemiluminescence reaction. The biosensor was tested for sensitivity, specificity, and overall practicality. Our results suggest that the detection of anti-E2 antibodies with this microbiosensor may enhance significantly HCV serological standard testing especially among patients during dialysis, which were diagnosed as HCV negative, by standard immunological tests, but were known to carry the virus. If transformed into an easy to use procedure, this assay might be used in the future as an important clinical tool for HCV screening in blood banks.

We demonstrate that it is possible to create surface-conductive fiber optics, upon which may be electropolymerized a biotinylated polypyrrole thin film, which may then be used to affinity coat the fiber with molecular recognition probes. This fiber-optic electroconductive surface modification is done by the deposition of a thin layer of indium tin oxide. Thereafter, biotin-pyrrole monomers are electropolymerized onto the conductive metal oxide surface and then exposed to avidin. Avidin-biotin interactions were used to modify the fiber optics with biotin-conjugated cholera toxin B subunit molecules, for the construction of an immunosensor to detect cholera antitoxin antibodies. The biosensor was tested for sensitivity, nonspecificity, and overall practicality.

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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.

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We report an innovative and useful procedure for immobilization of antibodies on to a fiber-optic silica surface. The procedure consists in the chemical oxidation of pyrrole-biotin monomers that are readily deposited as a thin film of poly(pyrrole-biotin) polymer on to the end-face of the fiber. The film was shown to be sufficiently translucent to enable photon coupling within the fiber transducer and its presence was demonstrated by means of fluorescent micrographs of bound rhodamine-labeled avidin. Fiber-optics modified with cholera toxin B subunit molecules were tested for sensitivity, non-specificity, and overall practicality. It was shown that the fiber-optic immuno-assay for the detection of anti-cholera toxin antibody was up to three orders of magnitude more sensitive than the classical enzyme-linked immunosorbent assay (ELISA).