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Nanoparticle–Protein Interactions: A Thermodynamic and Kinetic Study of the Adsorption of Bovine Serum Albumin to Gold Nanoparticle Surfaces
journal contribution
posted on 2013-12-03, 00:00 authored by Stefano
P. Boulos, Tyler A. Davis, Jie An Yang, Samuel E. Lohse, Alaaldin M. Alkilany, Lisa A. Holland, Catherine J. MurphyInvestigating the adsorption process
of proteins on nanoparticle
surfaces is essential to understand how to control the biological
interactions of functionalized nanoparticles. In this work, a library
of spherical and rod-shaped gold nanoparticles (GNPs) was used to
evaluate the process of protein adsorption to their surfaces. The
binding of a model protein (bovine serum albumin, BSA) to GNPs as
a function of particle shape, size, and surface charge was investigated.
Two independent comparative analytical methods were used to evaluate
the adsorption process: steady-state fluorescence quenching titration
and affinity capillary electrophoresis (ACE). Although under favorable
electrostatic conditions kinetic analysis showed a faster adsorption
of BSA to the surface of cationic GNPs, equilibrium binding constant
determinations indicated that BSA has a comparable binding affinity
to all of the GNPs tested, regardless of surface charge. BSA was even
found to adsorb strongly to GNPs with a pegylated/neutral surface.
However, these fluorescence titrations suffer from significant interference
from the strong light absorption of the GNPs. The BSA–GNP equilibrium
binding constants, as determined by the ACE method, were 105 times lower than values determined using spectroscopic titrations.
While both analytical methods could be suitable to determine the binding
constants for protein adsorption to NP surfaces, both methods have
limitations that complicate the determination of protein–GNP
binding constants. The optical properties of GNPs interfere with Ka determinations by static fluorescence quenching
analysis. ACE, in contrast, suffers from material compatibility issues,
as positively charged GNPs adhere to the walls of the capillary during
analysis. Researchers seeking to determine equilibrium binding constants
for protein–GNP interactions should therefore utilize as many
orthogonal techniques as possible to study a protein–GNP system.
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Keywords
nanoparticleBovine Serum Albuminmethodsurface chargeequilibrium binding constantsACEaffinity capillary electrophoresisadsorption processfluorescence quenching analysisNPfluorescence quenching titrationBSAGold Nanoparticle SurfacesInvestigatingdeterminationGNPmaterial compatibility issuesprotein adsorption
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