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Capillary waveguide fluoroimmunosensor with improved repeatability and detection sensitivity

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Abstract

An optical capillary waveguide fluoroimmunosensor based on glass capillaries internally coated with an ultrathin poly(dimethylsiloxane) (PDMS) film is presented. The evaluation of the capillaries developed was done in comparison with aminosilanized [3-(aminopropyl)triethoxysilane, APTES] glass and poly(methylpentene) (PMP) capillaries by immobilizing rabbit γ-globulins on the internal capillary wall. Following reaction with (R)-phycoerythrin-labelled antibody, the capillary was scanned with a laser beam and the fluorescence waveguided through the capillary wall was detected by a photomultiplier placed at one of its ends. The capillaries developed provided considerably improved protein coating homogeneity (intracapillary coefficients of variation 2.9–6.6%) and repeatability (intercapillary coefficients of variation 2.1–5.0%) compared with APTES-treated ones (7.9–13.4 and 8.5–15.2%, respectively). With use of these capillaries in a sandwich-type immunosensor for the determination of rabbit γ-globulins, the assay detection limit was improved eightfold (4.4 ng/mL) compared with that obtained using PMP capillaries (35.3 ng/mL), whereas the assay repeatability was improved threefold (intra-assay coefficients of variation 5.9–13.1%) compared with APTES-treated capillaries (15.6–36%).

Optoelectronic set-up used to scan the capillaries (left) and representative fluorescence scannings of dual-band poly(methylpentene) (PMP), PDMS-modified glass and APTES treated glass capillaries

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Acknowledgements

This work was partly supported by the BRITE/EURAM III project BOEMIS (BRPR CT-97-0393) and the Greek General Secretary for Research and Technology (GSRT) Program, I/R-RP-Excellence in Research II, 2005 (75% funded by the EU).

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Authors and Affiliations

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

    Aristidis E. Niotis, Christos Mastichiadis, Panagiota S. Petrou, Ion Christofidis & Sotirios E. Kakabakos

  2. Biochemistry Lab, Department of Chemistry, University of Athens, Panepistimiopolis, 15771, Athens, Greece

    Athanasia Siafaka-Kapadai

  3. Institute of Microelectronics, N.C.S.R. “Demokritos”, Aghia Paraskevi, 15310, Athens, Greece

    Konstantinos Misiakos

Authors
  1. Aristidis E. Niotis
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  2. Christos Mastichiadis
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  3. Panagiota S. Petrou
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  4. Ion Christofidis
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  5. Athanasia Siafaka-Kapadai
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  6. Konstantinos Misiakos
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  7. Sotirios E. Kakabakos
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Correspondence to Sotirios E. Kakabakos.

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Niotis, A.E., Mastichiadis, C., Petrou, P.S. et al. Capillary waveguide fluoroimmunosensor with improved repeatability and detection sensitivity. Anal Bioanal Chem 393, 1081–1086 (2009). https://doi.org/10.1007/s00216-008-2501-2

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  • DOI: https://doi.org/10.1007/s00216-008-2501-2

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