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Applying Total Internal Reflection Excitation and Super Critical Angle Fluorescence Detection to a Morphine Assay

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Abstract

A surface-sensitive fluorescence measurement platform is utilised in the detection of morphine. The platform is based on a polystyrene parabolic lens that enables the simultaneous application of total internal reflection excitation and supercritical angle fluorescence detection in the measurements. The molecular recognition of morphine is based on two antibodies, one against morphine and the other against the immune complex formed between the anti-morphine antibody and a morphine molecule. The antibodies are applied in a sandwich-like format in a one-step test, where the molecular binding onto a liquid-solid-interface is monitored in real time. Morphine concentrations between 0.6 and 18.2 ng/mL were reliably determined in 60 s, while concentrations down to 2.7 ng/mL were detected already in 20 s. With appropriate recognition molecules the technique is applicable also to other drugs and small analytes.

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References

  1. Kerrigan S, Phillips WH (2001) Comparison of ELISAs for drugs-of-abuse testing. Jn Clin Chem 47:540–547

    CAS  Google Scholar 

  2. Pulli T, Höyhtyä M, Söderlund H, Takkinen K (2005) One-step homogeneous immunoassay for small analytes. Anal Chem 77:2637–2642

    Article  CAS  PubMed  Google Scholar 

  3. Dams R, Choo RE, Lambert WE, Jones H, Huestis MA (2007) Oral fluid as an alternative matrix to monitor opiate and cocaine use in substance-abuse treatment patients. Drug Alcohol Depend 87:258–267

    Article  CAS  PubMed  Google Scholar 

  4. Välimäki H, Tappura K (2009) A novel platform for highly surface-sensitive fluorescent measurements applying simultaneous total internal reflection excitation and super critical angle detection. Chem Phys Lett 473:358–362

    Article  Google Scholar 

  5. Ruckstuhl T, Rankl M, Seeger S (2003) Highly sensitive biosensing using a supercritical angle fluorescence (SAF) instrument. Biosens Bioelectron 18:1193–1199

    Article  CAS  PubMed  Google Scholar 

  6. Enderlein J, Ruckstuhl T, Seeger S (1999) Highly efficient optical detection of surface-generated fluorescence. Appl Opt 38:724–732

    Article  CAS  PubMed  Google Scholar 

  7. Lakowicz JR (1999) Principles of fluorescence spectroscopy, 2nd edn. Kluwer Academic/Plenum Publishers, New York

    Google Scholar 

  8. O’Shannessy DJ, Brigham-Burke M, Soneson K, Hensley P, Brooks I (1993) Determination of rate equilibrium binding constants for macromolecular interactions using surface plasmon resonance: use of nonlinear least squares analysis Methods. Biochem Anal 212:457–468

    Article  Google Scholar 

  9. Vikholm-Lundin I (2005) Immunosensing based on site-directed immobilization of antibody fragments and polymers that reduce nonspecific binding. Langmuir 21:6473–6477

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Armi Boman for skilful technical assistance, Dr. Tarja Nevanen for commenting on the manuscript, and TEKES, the Finnish Funding Agency for Technology and Innovation, for financial support (40240/07).

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Authors and Affiliations

  1. VTT Technical Research Centre of Finland, P.O.Box 1300, FI-33101, Tampere, Finland

    Hannu S. Välimäki & Kirsi Tappura

  2. VTT Technical Research Centre of Finland, P.O.Box 1000, FI-02044, Espoo, Finland

    Timo Pulli

Authors
  1. Hannu S. Välimäki
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  2. Timo Pulli
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  3. Kirsi Tappura
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Correspondence to Hannu S. Välimäki.

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Välimäki, H.S., Pulli, T. & Tappura, K. Applying Total Internal Reflection Excitation and Super Critical Angle Fluorescence Detection to a Morphine Assay. J Fluoresc 20, 1003–1008 (2010). https://doi.org/10.1007/s10895-010-0647-8

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  • DOI: https://doi.org/10.1007/s10895-010-0647-8

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