Reversible versus irreversible binding to transferrin to polystyrene nanoparticles: Soft and hard corona

Silvia Milani, Francesca Baldelli Bombelli, Andrzej S. Pitek, Kenneth A. Dawson, Joachim Rädler

Research output: Contribution to journalArticlepeer-review

408 Citations (Scopus)


Protein adsorption to nanoparticles (NPs) is a key prerequisite to understand NP–cell interactions. While the layer thickness of the protein corona has been well characterized in many cases, the absolute number of bound proteins and their exchange dynamics in body fluids is difficult to assess. Here we measure the number of molecules adsorbed to sulfonate (PSOSO3H) and carboxyl-(PSCOOH) polystyrene NPs using fluorescence correlation spectroscopy. We find that the fraction of molecules bound to NPs falls onto a single, universal adsorption curve, if plotted as a function of molar protein-to-NP ratio. The adsorption curve shows the build-up of a strongly bound monolayer up to the point of monolayer saturation (at a geometrically defined protein-to-NP ratio), beyond which a secondary, weakly bound layer is formed. While the first layer is irreversibly bound (hard corona), the secondary layer (soft corona) exhibits dynamic exchange, if competing unlabeled is added. In the presence of plasma proteins, the hard corona is stable, while the soft corona is almost completely removed. The existence of two distinct time scales in the protein off-kinetics, for both NP types studied here, indicates the possibility of an exposure memory effect in the NP corona.
Original languageEnglish
Pages (from-to)2532-2541
Number of pages10
JournalACS Nano
Issue number3
Early online date22 Feb 2012
Publication statusPublished - 2012


  • fluorescence correlation spectroscopy (FCS)
  • nanoparticles−proteins interaction
  • protein corona
  • bionano interface
  • Transferrin

Cite this