July 14-17, 2019
This work presents a novel technique to characterize the surface hydrophobicity and other surface properties of proteins. The surface properties of industrial and therapeutic proteins are key to understanding their behavior 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 measurements of the surface hydrophobicity of column resin-bound proteins using changes in probe retention behavior. The dimensionless hydrophobicity factor (Hf) for Lysozyme and BSA was obtained with changing pH and temperature. The two sets of results followed published fluorescence spectroscopy data, where available, or hypotheses based on other data in the literature. The effects of pH appeared to be reversible. The effects of temperature were not reversible within the given experimental setup. Some interesting observations were made on protein unfolding and conformation. This new technique could provide reliable and quick surface parameter data sets using standard equipment and methods for any conceivable protein of interest.
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Dilip Sethi, Sarah Hedberg, and Daryl Williams, "A novel technique to characterize the surface hydrophobicity of proteins using inverse liquid chromatography" in "Biological and Pharmaceutical Complex Fluids III: Protein Self-Assembly, Rheology and Interfacial Properties", Samiul Amin, Manhattan College, USA Miguel Rodrigues, University of Lisbon, Portugal Paolo Arosio, ETHZ, Switzerland Eds, ECI Symposium Series, (2019). https://dc.engconfintl.org/bpcf_iii/13