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Capillary waveguide fluoroimmunosensor with improved repeatability and detection sensitivity.

Aristidis E Niotis1, Christos Mastichiadis, Panagiota S Petrou

  • 1Immunoassay-Immunosensors Laboratory, Institute of Radioisotopes & Radiodiagnostic Products, N.C.S.R. Demokritos, Aghia Paraskevi, 15310, Athens, Greece.

Analytical and Bioanalytical Chemistry
|November 13, 2008
PubMed
Summary
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This study introduces a novel optical capillary waveguide fluoroimmunosensor using poly(dimethylsiloxane) (PDMS) coatings. The new sensor significantly improves protein coating, detection limits, and repeatability for detecting rabbit gamma-globulins.

Area of Science:

  • Biomedical Engineering
  • Analytical Chemistry
  • Materials Science

Background:

  • Traditional fluoroimmunosensors face challenges with protein coating homogeneity and assay sensitivity.
  • Glass and poly(methylpentene) capillaries have limitations in surface modification for biosensing applications.

Purpose of the Study:

  • To develop and evaluate an optical capillary waveguide fluoroimmunosensor utilizing ultrathin poly(dimethylsiloxane) (PDMS) films.
  • To compare the performance of PDMS-coated capillaries with aminosilanized (APTES) and poly(methylpentene) (PMP) capillaries for immunoassay applications.

Main Methods:

  • Fabrication of glass capillaries internally coated with ultrathin PDMS films.
  • Immobilization of rabbit gamma-globulins and subsequent reaction with fluorescently labeled antibodies.

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  • Fluorescence detection using laser scanning and a photomultiplier for signal acquisition.
  • Comparative analysis of protein coating homogeneity, repeatability, and assay performance.

    • Main Results:

    • PDMS-coated capillaries demonstrated significantly improved protein coating homogeneity (CV 2.9-6.6%) and repeatability (CV 2.1-5.0%) compared to APTES-treated capillaries (CV 7.9-13.4% and 8.5-15.2%).
    • The fluoroimmunosensor using PDMS capillaries achieved an eightfold improvement in detection limit (4.4 ng/mL) compared to PMP capillaries (35.3 ng/mL).
    • Assay repeatability was threefold improved (CV 5.9-13.1%) with PDMS capillaries compared to APTES-treated capillaries (CV 15.6-36%).

    Conclusions:

    • Ultrathin PDMS coatings on glass capillaries offer superior performance for fluoroimmunosensor development.
    • The developed optical capillary waveguide fluoroimmunosensor provides enhanced sensitivity and repeatability for protein detection.
    • This technology holds promise for improved diagnostic assays and biosensing applications.