(548c) A Novel Technique to Characterise the Surface Hydrophobicity of Proteins Using Inverse Liquid Chromatography
AIChE Annual Meeting
2019
2019 AIChE Annual Meeting
Food, Pharmaceutical & Bioengineering Division
Protein Science and Engineering: Advances on Engineering the Physiochemical Properties of Proteins
Wednesday, November 13, 2019 - 1:06pm to 1:24pm
The surface properties of industrial and therapeutic proteins are key to understanding their behaviour in-situ, in the laboratory and in processes. Protein surface hydrophobicity is a marker for three dimensional structure, stability and function. Inverse Liquid Chromatography of Proteins (ILCP) with small molecule hydrophobic probes can be used to obtain direct thermodynamic measurements of the surface hydrophobicity of column resin-bound proteins using changes in probe retention behaviour. The dimensionless hydrophobicity factor (Hf) for Lysozyme and BSA was obtained with changing pH and temperature. The two sets of results are similar to published data from fluorescence and dielectric spectroscopy. The effects of pH appear to be reversible. The effects of temperature are reversible if the thermodynamic load is mild. This novel technique could provide reliable and quick hydrophobicity and other surface parameter data sets using standard equipment and methods for any conceivable protein of interest.