Component‐Specific Analysis of Plasma Protein Corona Formation on Gold Nanoparticles Using Multiplexed Surface Plasmon Resonance

Abstract

At the nano-bio interface, human plasma differentially interacts with engineered nanomaterials through the creation of protein coronas, which in turn become primary determinants of both the pharmacokinetics and pharmacodynamics of circulating nanoparticles. Here, for the first time, the specific binding kinetics of the four major corona forming proteins (human serum albumin, fibrinogen, ApoA1, and polyclonal IgG) are determined for gold nanoparticles (AuNPs). Using a multiplexed surface plasmonic assay, highly reproducible measurements of on rate (k(on)), off rate (k(off)), and disassociation constant (K(D)), in addition to relative amounts of protein binding, are obtained. Dramatic differences in k(on) for individual components are shown as primary determinants of protein affinities, with k(on) ranging over nearly two orders of magnitude for the proteins studied, while k(off) remains within a factor of two for the set. The effect of polyethylene glycol (PEG) modification on plasma component binding is also studied and the effect of PEG length on human serum interaction is characterized through systematic screening of PEG molecular weight (2-30k). The effect of nanoparticle modification on particle targeting is also characterized through study of a hybrid AuNP system.

Techniques for the experimental investigation of the protein corona

“…The binding of proteins to NPs can be detected as a change in the angular position of the SPR peak. Multiplexed SPR using microchips with multiple microfluidic channels has been used for the simultaneous analysis of the interaction of Au NPs with several plasma components [66]. Also second harmonic light scattering (SHLS) has been proposed as a powerful technique for the quantification of weakly interacting proteins, which cannot be easily detected by conventional techniques [67].…”

Effect of the Protein Corona on Antibody–Antigen Binding in Nanoparticle Sandwich Immunoassays

“…1 Protein coronas mediate the interactions between NPs and their environment, influencing processes such as association with cell surfaces and intracellular uptake. 2–6 Often protein coronas have negative impacts on these processes, as the corona around the NP can actually obscure its targeting ligand, reducing the specificity of interaction of the targeting species on the NP surface. 7–9 The impact of the protein corona has been studied predominantly for aspects of nanomedicine concerned with the transport, blood circulation, and cell interactions of nanomaterials such as targeting and cell uptake.…”

Biological Behavior Regulation of Gold Nanoparticles via the Protein Corona

“…These proteins already existing on the GNP surface dynamically compete with other proteins that have slower association rates but eventually higher binding affinities. [ 28a ] With the assistance of the analytical techniques mentioned above, the nature of the PC on the surface of GNPs mainly depends on i) the specific characteristics of the biological environment in which GNPs are dispersed and ii) the physicochemical features of the GNPs, including the size, shape, and surface chemistry.…”

“…For example, when the shape of GNPs shifts from spherical to triangular, the helicities of HB and HSA adsorbed on GNPs were reduced. [ 28b ] The amount of proteins adsorbed on the surface of GNPs (11 nm) was reported to be an order of magnitude lower than that on the surface of GNRs (10 nm × 36 nm). This significant difference lies in the different surface areas of the spherical GNPs (530 nm 2 ) and club‐shaped GNRs (1550 nm 2 ).…”