Techniques for the experimental investigation of the protein corona

Abstract

Due to its enormous relevance the corona formation of adsorbed proteins around nanoparticles is widely investigated. A comparison of different experimental techniques is given. Direct measurements of proteins, such as typically performed with mass spectrometry, will be compared with indirect analysis, in which instead information about the protein corona is gathered from changes in the properties of the nanoparticles. The type of measurement determines also whether before analysis purification from unbound excess proteins is necessary, which may change the equilibrium, or if measurements can be performed in situ without required purification. Pros and contras of the different methods will be discussed.

Super‐Resolution Microscopy Unveils Dynamic Heterogeneities in Nanoparticle Protein Corona

“…[6][7][8][9] The approaches being used to study the corona phenomenon can be divided in ex situ and in situ techniques, and mainly study the structure of the protein corona, the affinity, the conformation, and the composition. [6,10,11] However, the small size of nanoparticles together with the complexity of serum composition makes challenging the study of nanoparticle-protein interaction. [3] Moreover, the corona is believed to be dynamic, changing in composition in time.…”

“…Therefore, it is of major importance to understand the formation and the dynamics of nanoparticle protein corona, permitting the design of effective nanostructures for biomedical applications . The approaches being used to study the corona phenomenon can be divided in ex situ and in situ techniques, and mainly study the structure of the protein corona, the affinity, the conformation, and the composition . However, the small size of nanoparticles together with the complexity of serum composition makes challenging the study of nanoparticle–protein interaction .…”

Formation of a Monolayer Protein Corona around Polystyrene Nanoparticles and Implications for Nanoparticle Agglomeration

“…Weak binding results in a “soft corona,” characterized by fast dynamic exchange between adsorbed proteins and those in solution, whereas a tightly bound, “hard corona” is essentially persistent . Technically, hard coronae are easier to characterize because the NPs can be separated from the biofluid together with the adsorbed protein layer . Major structural changes of the adsorbed proteins up to complete denaturation may occur during hard corona formation .…”

An apolipoprotein-enriched biomolecular corona switches the cellular uptake mechanism and trafficking pathway of lipid nanoparticles

“…It is precisely this new identity, usually referred to as “biological identity”, which controls the biological fate of NPs, including their circulation time, immune system activation, accumulation in liver and spleen, removal from the bloodstream via the mononuclear phagocytic system (MPS), biodistribution, and cellular interactions 7 . Concerning cellular interactions in particular, since the function of nanomaterials is expected to be carried out in a specific cellular location, 10, 11 accurate characterization of BC 12 and understanding NP-BC-cell interactions is a critical step to design safer and more efficient NPs as well as to predict possible toxicity effects 13–16 . After cellular association, most nanomaterials get internalized via endocytic pathways 17–19 .…”