Biomedical Engineering Reference
In-Depth Information
Table 4.1
Overview of analytical strategies to monitor NP-protein or NP-proteome interactions (Some of the data collected from [
3
,
4
])
Studies
Analysis methods
Mechanism
Advantages
Limitations
Comment
Binding affinity
and ratio
UV-visible
spectroscopy
(UV-Vis)
Change in light absorption
spectra or plasmonic
absorption due to
adsorption of protein
on NP
Fast, flexible, simple
sample preparation,
qualitative method
Absorption spectrum is
dependent on NP type,
pH, solvent, interfering
substances, and light
scattering of NP
For quantitative and
conclusive results, it
should be combined with
other techniques
Fluorescence
spectroscopy
(FS)
Absorption of light by
fluorophores and
fluorescence emission
Sensitive, quantitative
Protein or NP should be
fluorescent or labeled
with fluorophores
Labeling with fluorophores
can change the surface
properties and protein
corona structure
Isothermal titration
calorimetry
(ITC)
The change in temperature
upon addition of proteins
to NPs is recorded
Binding affinity and
binding
stoichiometry can
be measured
Dynamic Light
Scattering (DLS)
Size distribution
Only for spherical NP
Atomic Force
Microscopy
(AFM)
3D surface profile
Limited scanning area
Conformational
changes of
NP-bound
proteins
Circular dichroism
(CD)
Interaction of left- and right-
handed circularly
polarized light with
chiral structures
Secondary structures
of proteins
No residue-specific
information, for single
proteins
Due to the high noise in the
far UV (
200 nm),
calculation of second
structure in this region is
not accurate
<
Fourier transform
infrared
spectroscopy
(FTIR)
Objects have to be dried
