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Effects of immobilization on a FRET immunosensor for the detection of myocardial infarction

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Abstract

A novel optical biosensor technique is being developed for the early detection of myocardial infarction by utilizing the distance-dependent chemical transduction method of fluorescence resonance energy transfer (FRET). The FRET process requires two fluorophores termed the donor and the acceptor. When in close proximity, the donor absorbs energy from the excitation source and non-radiatively transfers the energy to the acceptor, which in turn emits fluorescent energy. This distance-dependent property was utilized to detect conformational changes when antibodies combine with their respective antigens. The fluorophores were conjugated to an antibody–Protein A complex and then immobilized via silanization to the distal ends of optical fibers. Three different antibody–Protein A complexes were immobilized: generic IgG, cardiac Troponin T (cTnT), and cardiac Troponin I (cTnI). Results showed that upon the addition of the specific antigens, the antibodies underwent a conformational change, reducing the distance between the FRET fluorophores. The generic IgG responded to 233 nM antigens, whereas the cTnT biosensor had a limit of detection of 75 nM, and the cTnI biosensors had a limit of detection of 94 nM.

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Acknowledgements

This work was supported, in part, by an Industrial Grant from Medtronic, Inc. (Fridley, MN), by the Missouri Agricultural Experiment Station, and the Missouri Food for the 21st Century.

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Authors and Affiliations

  1. Department of Biological Engineering, University of Missouri—Columbia, Columbia, MO 65211, USA

    Sheila A Grant, Mary E Pierce & Darcy J Lichlyter

  2. Department of Electrical and Computer Engineering, University of Missouri—Columbia, Columbia, MO 65211, USA

    David A Grant

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  1. Sheila A Grant
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  2. Mary E Pierce
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  3. Darcy J Lichlyter
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  4. David A Grant
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Correspondence to Sheila A Grant.

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Grant, S.A., Pierce, M.E., Lichlyter, D.J. et al. Effects of immobilization on a FRET immunosensor for the detection of myocardial infarction. Anal Bioanal Chem 381, 1012–1018 (2005). https://doi.org/10.1007/s00216-004-2976-4

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  • DOI: https://doi.org/10.1007/s00216-004-2976-4

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