“…Recent progress in the field has allowed the inclusion of cryo-probes and the development of electron crystallography (used to determine the structure of several proteins allowing the identification, in some cases, of secondary structure motifs) [97, 98]. Other techniques [91] used to support research related to protein adsorption include capillary electrophoresis [99], mass spectrometry [100, 101] and particle mass spectrometry [102] (to identify and quantify proteins through fragmentation), various vibrational spectroscopictechniques [103, 104] (to study protein/substrate interface and elucidate the biosensing mechanism), UV-Vis spectroscopy [105], Fourier-transform infrared (FTIR) spectroscopy[106, 107] (to analyze sequential and competitive adsorption), circular dichroism (CD) spectroscopy[108, 109] (to investigate the secondary structure of adsorbed proteins), isothermal titration calorimetry [110] (to investigate thermodynamic properties of interactions), surface plasmon resonance (SPR)[37, 111-113] (for quantitation of proteins and kinetic studies but limited to gold or silver surfaces), total internal reflection fluorescence spectroscopy [114] (for determining conformational changes in adsorbed proteins), infrared spectroscopy [115] (to observe protein conformation and load but with limited use in multicomponent protein solutions because of complex spectra), and fluorescence spectroscopy[84, 116] (to observe structural change in protein and protein loading).As their interaction with proteins can lead to spectral shifts in gold nanoparticles(AuNP), various colorimetric assays have been presented to monitor either protein-protein interactions [117] or conformational changes [118]. Additionally, it was recently reported that in the low protein coverage regime, nanoparticle tracking analysis, and differential dynamic light scattering (but not differential centrifugal sedimentation)correlate with the expected plasmon frequency shift of AuNP[119].…”