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A new optical platform for biosensing based on fluorescence anisotropy

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Abstract

A novel fluorescence-based optical platform for the interrogation of an optical biochip was designed and developed. The optical biochip was made of poly(methyl methacrylate) (PMMA) formed by two pieces of PMMA appropriately shaped in order to obtain four microchannels that are 500-μm wide and 400-μm high. The lower part includes the microchannels and the inlet and outlet for the fluidics, while the sensing biolayer was immobilized on the upper part. The optical signal comprised the fluorescence emitted by the biolayer, which was anisotropically coupled to the PMMA cover and suitably guided by the PMMA chip. The potentiality of the optical chip as a biosensor was investigated by means of a direct IgG/anti-IgG interaction carried out inside the flow channels. The mouse-IgG was covalently immobilized on the internal wall of the PMMA cover, and the Cy5-labelled anti-mouse IgG was used for the specific interaction. Several chemical treatments of the PMMA surface were investigated, poly(L-lactic acid), Eudragit L100 and NaOH, in order to obtain the most effective distribution of carboxylic groups useful for the covalent immobilisation of the mouse-IgG. The treatment with Eudragit L100 was found to be the most successful. Limits of detection and quantification of 0.05 μg mL−1 and 0.2 μg mL−1, respectively, were obtained with the configuration described.

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Acknowledgments

This research study was supported by the European Community within the framework of the five-year EU funded project under IST priority, CAREMAN (HealthCARE by Biosensor Measurements and Networking). The authors wish to thank Lorena Tedeschi of the Institute of Clinical Physiology of Pisa for her positive collaboration and fruitful suggestions.

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Authors and Affiliations

  1. Institute of Applied Physics, CNR, Via Madonna del Piano 10, 50019, Sesto Fiorentino (FI), Italy

    F. Baldini, A. Carloni, A. Giannetti & C. Trono

  2. Datamed S.r.L., Via Papa Giovanni XXIII 45, 20090, Rodano (MI), Italy

    G. Porro

Authors
  1. F. Baldini
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  2. A. Carloni
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  3. A. Giannetti
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  4. G. Porro
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  5. C. Trono
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Correspondence to F. Baldini.

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Baldini, F., Carloni, A., Giannetti, A. et al. A new optical platform for biosensing based on fluorescence anisotropy. Anal Bioanal Chem 391, 1837–1844 (2008). https://doi.org/10.1007/s00216-008-1904-4

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  • DOI: https://doi.org/10.1007/s00216-008-1904-4

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