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Synthesis of polystyrene microspheres and functionalization with Pd0 nanoparticles to perform bioorthogonal organometallic chemistry in living cells

Nature Protocols volume 7, pages 1207–1218 (2012)Cite this article

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Abstract

We have developed miniaturized heterogeneous Pd0-catalysts (Pd0-microspheres) with the ability to enter cells, stay harmlessly within the cytosol and mediate efficient bioorthogonal organometallic chemistries (e.g., allylcarbamate cleavage and Suzuki-Miyaura cross-coupling). This approach is a major addition to the toolbox available for performing chemical reactions within cells. Here we describe a full protocol for the synthesis of the Pd0-microspheres from readily available starting materials (by the synthesis of size-controlled amino-functionalized polystyrene microspheres), as well as for their characterization (electron microscopy and palladium quantitation) and functional validation ('in solution' and 'in cytoplasm' conversions). From the beginning of the synthesis to functional evaluation of the catalytic device requires 5 d of work.

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Figure 1
Figure 2
Figure 3: Physical characterization of microspheres.
Figure 4: TEM images of 0.5-μm microspheres.
Figure 5: Functional validation study of Pd0 microspheres.

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Acknowledgements

This work was supported financially by the Engineering and Physical Sciences Research Council (M.B.), the Swiss National Science Foundation (E.M.V.J.) and the Government of Malaysia (R.M.Y.). A.U.-B. and R.M.S.-M. thank the Medical Research Council Institute of Genetics and Molecular Medicine and the Royal Society, respectively, for funding. We are grateful to F. Thielbeer and J. Cardenas-Maestre for their helpful advice regarding the microsphere synthesis and to J. Weiss for testing the protocol.

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Author notes

  1. Asier Unciti-Broceta, Emma M V Johansson and Rahimi M Yusop: These authors contributed equally to this work.

Authors and Affiliations

  1. Edinburgh Cancer Research UK Centre, Medical Research Council Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UK

    Asier Unciti-Broceta

  2. School of Chemistry, University of Edinburgh, Edinburgh, UK

    Emma M V Johansson, Rahimi M Yusop & Mark Bradley

  3. Faculty of Science and Technology, School of Chemical Sciences and Food Technology, Universiti Kebangsaan Malaysia, Selangor Darul Ehsan, Malaysia

    Rahimi M Yusop

  4. Facultad de Farmacia, Universidad de Granada, Campus de la Cartuja, Granada, Spain

    Rosario M Sánchez-Martín

Authors
  1. Asier Unciti-Broceta
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  2. Emma M V Johansson
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  3. Rahimi M Yusop
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  4. Rosario M Sánchez-Martín
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  5. Mark Bradley
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Contributions

A.U.-B. designed, supervised and wrote the protocol; E.M.V.J. developed and wrote the protocol; R.M.Y. developed the protocol; R.M.S.-M. developed the protocol; M.B. designed, supervised and wrote the protocol.

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Correspondence to Asier Unciti-Broceta or Mark Bradley.

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The authors declare no competing financial interests.

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Unciti-Broceta, A., Johansson, E., Yusop, R. et al. Synthesis of polystyrene microspheres and functionalization with Pd0 nanoparticles to perform bioorthogonal organometallic chemistry in living cells. Nat Protoc 7, 1207–1218 (2012). https://doi.org/10.1038/nprot.2012.052

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