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Chemosensors and biosensors based on polyelectrolyte microcapsules containing fluorescent dyes and enzymes

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Abstract

The concept of enzyme-assisted substrate sensing based on use of fluorescent markers to detect the products of enzymatic reaction has been investigated by fabrication of micron-scale polyelectrolyte capsules containing enzymes and dyes in one entity. Microcapsules approximately 5 μm in size entrap glucose oxidase or lactate oxidase, with peroxidase, together with the corresponding markers Tris(4,7-diphenyl-1,10-phenanthroline)ruthenium(II) dichloride (Ru(dpp)) complex and dihydrorhodamine 123 (DHR123), which are sensitive to oxygen and hydrogen peroxide, respectively. These capsules are produced by co-precipitation of calcium carbonate particles with the enzyme followed by layer-by-layer assembly of polyelectrolytes over the surface of the particles and incorporation of the dye in the capsule interior or in the multilayer shell. After dissolution of the calcium carbonate the enzymes and dyes remain in the multilayer capsules. In this study we produced enzyme-containing microcapsules sensitive to glucose and lactate. Calibration curves based on fluorescence intensity of Ru(dpp) and DHR123 were linearly dependent on substrate concentration, enabling reliable sensing in the millimolar range. The main advantages of using these capsules with optical recording is the possibility of building single capsule-based sensors. The response from individual capsules was observed by confocal microscopy as increasing fluorescence intensity of the capsule on addition of lactate at millimolar concentrations. Because internalization of the micron-sized multi-component capsules was feasible, they could be further optimized for in-situ intracellular sensing and metabolite monitoring on the basis of fluorescence reporting.

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Acknowledgements

L.K. is grateful for a DAAD stipend. The authors are grateful to Professor Helmuth Möhwald for continuous support and stimulating discussion. This project was funded by Ministry of Education and Science of the Russian Federation, contracts 02.740.11.5226 and 14.740.11.1363.

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Authors and Affiliations

  1. Institute of Theoretical and Experimental Biophysics RAS, Institutskaya street 3, 142290, Pushchino, Moscow Region, Russia

    Lyubov I. Kazakova, Lyudmila I. Shabarchina & Gleb B. Sukhorukov

  2. School of Engineering and Materials Science, Queen Mary University of London, Mile End Road, E1 4NS, London, UK

    Salzitsa Anastasova, Anton M. Pavlov, Pankaj Vadgama & Gleb B. Sukhorukov

  3. Max Planck Institute of Colloids and Interface, am Muhlenberg 1, 14424, Potsdam/Golm, Germany

    Lyubov I. Kazakova & Andre G. Skirtach

  4. Department of Molecular Biotechnology & NanoBio-Photonics, Ghent University, 9000, Ghent, Belgium

    Andre G. Skirtach

Authors
  1. Lyubov I. Kazakova
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  2. Lyudmila I. Shabarchina
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  3. Salzitsa Anastasova
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  4. Anton M. Pavlov
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  5. Pankaj Vadgama
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  6. Andre G. Skirtach
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  7. Gleb B. Sukhorukov
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Corresponding author

Correspondence to Gleb B. Sukhorukov.

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Published in the topical collection Characterization of Thin Films and Membranes with guest editors Daniel Mandler and Pankaj Vadgama.

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Kazakova, L.I., Shabarchina, L.I., Anastasova, S. et al. Chemosensors and biosensors based on polyelectrolyte microcapsules containing fluorescent dyes and enzymes. Anal Bioanal Chem 405, 1559–1568 (2013). https://doi.org/10.1007/s00216-012-6381-0

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  • DOI: https://doi.org/10.1007/s00216-012-6381-0

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