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Evaluation of the derivates of phosphorescent Pt-coproporphyrin as intracellular oxygen-sensitive probes

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Abstract

Several new derivatives of the phosphorescent Pt(II)-coproporphyrin (PtCP) were evaluated with respect to the sensing of intracellular oxygen by phosphorescence quenching. Despite the more favorable molecular charge compared to PtCP, self-loading into mammalian cells was rather inefficient for all the dyes, while cell loading by facilitated transport using transfection reagents produced promising results. The PtCP-NH2 derivative, which gave best loading efficiency and S/N ratio, was investigated in detail including the optimisation of loading conditions, studies of sub-cellular localization, cytotoxicity, oxygen sensitivity and long-term signal stability. Being spectrally similar to the macromolecular MitoXpress™ probe currently used in this application, the PtCP-NH2 demonstrated higher loading efficiency and phosphorescent signals, suitability for several problematic cell lines and a slightly increased lifetime scale for the physiological range (0–200 μM O2). In physiological experiments with different cell types, mitochondrial uncouplers and inhibitors performed on a time-resolved fluorescence plate reader, this probe produced the anticipated profiles of intracellular oxygen concentration and responses to cell stimulation. Therefore, PtCP-NH2 represents a convenient probe for the experiments and applications in which monitoring of cellular oxygen levels is required.

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Acknowledgments

This work was supported by the Science Foundation of Ireland, grant 07/IN.1/B1804. The authors are grateful to R. Dmitriev and A. Zhdanov for the help with PC12 cells, G. Jasionek for the help with HepG2 cell line, and Dr. J. Hynes for assistance with oxygen sensing experiments.

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Authors and Affiliations

  1. Biochemistry Department, Cavanagh Building, University College Cork, Cork, Ireland

    Andreas Fercher & Dmitri Papkovsky

  2. Institute of Biomedical Chemistry, Pogodinskaya Str. 10, 119121, Moscow, Russia

    Gelii V. Ponomarev & Dmitri Yashunski

  3. Biochemistry Department, University College Cork, Cavanagh Pharmacy Building, College Road, Cork, Ireland

    Dmitri Papkovsky

Authors
  1. Andreas Fercher
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  2. Gelii V. Ponomarev
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  3. Dmitri Yashunski
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  4. Dmitri Papkovsky
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Correspondence to Dmitri Papkovsky.

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Fercher, A., Ponomarev, G.V., Yashunski, D. et al. Evaluation of the derivates of phosphorescent Pt-coproporphyrin as intracellular oxygen-sensitive probes. Anal Bioanal Chem 396, 1793–1803 (2010). https://doi.org/10.1007/s00216-009-3399-z

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  • DOI: https://doi.org/10.1007/s00216-009-3399-z

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