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A quantitative fluorescence study of protein monolayer formation on colloidal nanoparticles

Nature Nanotechnology volume 4, pages 577–580 (2009)Cite this article

Abstract

It is now known that nanoparticles, when exposed to biological fluid, become coated with proteins and other biomolecules to form a ‘protein corona’1. Recent systematic studies have identified various proteins that can make up this corona, but these nanoparticle–protein interactions are still poorly understood, and quantitative studies to characterize them are few in number. Here, we have quantitatively analysed the adsorption of human serum albumin onto small (10–20 nm in diameter) polymer-coated FePt and CdSe/ZnS nanoparticles by using fluorescence correlation spectroscopy. The protein corona forms a monolayer with a thickness of 3.3 nm. Proteins bind to the negatively charged nanoparticles with micromolar affinity, and time-resolved fluorescence quenching experiments show that they reside on the particle for ∼100 s. These new findings deepen our quantitative understanding of the protein corona, which is of utmost importance in the safe application of nanoscale objects in living organisms.

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Figure 1: Structure of HSA and the protein corona.
Figure 2: Examples of FCS autocorrelation functions of polymer-coated FePt nanoparticles in the absence and presence of HSA.
Figure 3: Association of HSA to polymer-coated FePt nanoparticles measured by FCS.
Figure 4: Kinetic studies of HSA association to polymer-coated FePt nanoparticles.

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Acknowledgements

This work was supported by the Deutsche Forschungsgemeinschaft (DFG) through the Center for Functional Nanostructures (CFN) and Schwerpunktprogramm (SPP) 1313, grants NI291/7 and PA794/4.

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Authors and Affiliations

  1. Institute of Biophysics, University of Ulm, Albert Einstein-Allee 11, Ulm, 89081, Germany

    Carlheinz Röcker, Matthias Pötzl & G. Ulrich Nienhaus

  2. Fachbereich Physik, Philipps Universität Marburg, Renthof 7, Marburg, 35037, Germany

    Feng Zhang & Wolfgang J. Parak

  3. Institute of Applied Physics and Center for Functional Nanostructures, University of Karlsruhe (TH), Karlsruhe, 76128, Germany

    G. Ulrich Nienhaus

  4. Department of Physics, University of Illinois at Urbana-Champaign, Urbana, 61801, Illinois, USA

    G. Ulrich Nienhaus

Authors
  1. Carlheinz Röcker
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  2. Matthias Pötzl
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  3. Feng Zhang
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Contributions

C.R. and G.U.N. conceived and designed the experiments. C.R. and M.P. performed the experiments. C.R. and M.P. analysed the data. F.Z. and W.P. contributed materials. C.R., W.P. and G.U.N. co-wrote the paper. All authors discussed the results and commented on the manuscript.

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Correspondence to G. Ulrich Nienhaus.

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Röcker, C., Pötzl, M., Zhang, F. et al. A quantitative fluorescence study of protein monolayer formation on colloidal nanoparticles. Nature Nanotech 4, 577–580 (2009). https://doi.org/10.1038/nnano.2009.195

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