In situ monitoring of protein adsorption on a nanoparticulated gold film by attenuated total reflection surface-enhanced infrared absorption spectroscopy.
ABSTRACT In situ surface enhanced infrared absorption spectroscopy (SEIRAS) with an attenuated total reflection (ATR) configuration has been used to monitor the adsorption kinetics of bovine hemoglobin (BHb) on a Au nanoparticle (NP) film. The IR absorbance for BHb molecules on a gold nanoparticle film deposited on a Si hemispherical optical window is about 58 times higher than that on a bare Si optical window and the detection sensitivity has been improved by 3 orders of magnitude. From the IR signal as a function of adsorption time, the adsorption kinetics and thermodynamics can be explored in situ. It is found that both the electrostatic interaction and the coordination bonds between BHb residues and Au NP film surface affect the adsorption kinetics. The maximum adsorption can be obtained in solution pH 7.0 (close to the isoelectric point of the protein) due to the electrostatic interaction among proteins. In addition, the isotherm of BHb adsorption follows well the Freundlich adsorption model.
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ABSTRACT: With the advantages of in situ analysis and high surface sensitivity, surface-enhanced infrared absorption spectroscopy in attenuated total reflection mode (ATR-SEIRAS) combined with electrochemical methods has been employed to examine the interfacial direct electron transfer activity and adsorption kinetics of cytochrome c (cyt c). This work presents data on cyt c adsorption onto negatively charged mercaptohexanoic acid (MHA) and positively charged 6-amino-1-hexanethiol (MHN) self-assembled monolayers (SAMs) on gold nanofilm surfaces. The adsorbed cyt c displays a higher apparent electron transfer rate constant (33.5 ± 2.4 s(-1)) and apparent binding rate constant (73.1 ± 5.2 M(-1) s(-1)) at the MHA SAMs surface than those on the MHN SAMs surface. The results demonstrate that the surface charge density determines the protein adsorption kinetics, while the surface charge character determines the conformation and orientation of proteins assembled which in turn affects the direct electron transfer activity.The Analyst 08/2013; · 4.23 Impact Factor