Use of Fourier Transform Infrared/Attenuated Total Reflectance Spectroscopy for the Study of Surface Adsorption of Proteins

Abstract

The adsorption behavior of lysozyme and immunoglobulin G (IgG) onto a ZnSe crystal surface has been detected by the FT-IR/ATR technique. With this technique we are able to detect the protein adsorption process at a very low concentration (0.0005 mg/mL). The equation of Sperline et al. [Langmuir 3, 198 (1987)] was successfully applied to calculate the adsorption density of protein from an aqueous solution on a real-time basis. The monolayer formation of lysozyme was observed at a concentration range of 10⁻⁷ M to 10⁻⁵ M. A monolayer-to-multilayer adsorption transition of lysozyme was clearly observed from the adsorption isotherm plot at 35 μM. A differential adsorption density was observed for IgG that could be explained on the basis of its differential size and surface charge.

Keywords

ATR spectroscopy Infrared Spectroscopic techniques Adsorption density Amide II Surface Zinc selenide

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