Biomedical Engineering Reference
In-Depth Information
Chapter 4
Analytical Methods for Corona Evaluations
Abstract
In order to have deep understanding on the nature and composition of the
formed protein corona, one should have adequate information on the available
characterization techniques. In this chapter, comprehensive descriptions on the
protein corona evaluation methods (e.g., spectroscopy methods (UV/Vis, Raman,
fluorescence, mass spectrometry, nuclear magnetic resonance, etc.), dynamic light
scattering, circular dichroism, differential centrifugal sedimentation, scanning and
transmission electron microscopies, X-ray crystallography, chromatography, etc.)
together with their limitations are provided.
When nanoparticles (NPs) are in contact with biological fluids (serum, plasma,
etc.), they can strongly interact with proteins or other biomolecules that dramati-
cally change their surface proprieties. Their surface can acquire a new biological
identity which will influence the stability and the interaction with the living
material. Thus, the in vivo experiments of NP biodistribution can be affected
[
1
]. There is an important interest to develop analytical methods to investigate
NP-protein interactions. The study of interaction of proteins (determination of
binding rates, affinities, stoichiometries of protein association, etc.) with
nanoparticles in biological fluids is particularly complex since more than 3,700
proteins in different concentrations are coexisting and competing for binding to the
surface of the nanoparticle [
2
]. Li et al. [
3
] and Mahmoudi et al. [
4
] reviewed
characterization methods to study NP-protein interactions. These characterization
techniques are summarized and classified based on their application in NP-protein
studies in Table
4.1
. In this chapter each of these techniques will be discussed with
details. It should be mentioned that some of these techniques (such as UV-Vis, FS,
DLS, CD, FTIR, and ITC) are only employed to study interaction of a single protein
with NP, while other techniques such as chromatography, electrophoresis, MS,
SPR, and QCM can investigate the interaction of many available proteins (prote-
ome) in the environment with the NPs.
A conventional approach to study the NP corona involves incubation of NPs
with complex mixtures of proteins (e.g., plasma or serum) for various periods of
