Research Area: Characterisation of Protein Surfaces
Characterisation of Protein Surfaces using Inverse Liquid Chromatography
Protein surfaces hold the key to their behaviour in a natural, therapeutic and industrial environment. This research aims to use Inverse Liquid Chromatography (via HPLC) on immobilised proteins to characterise their, often dynamically changing, surfaces. This includes an exploration of hydrophobicity, charge, degradation, function and more, using appropriate, complementary small molecule probes.
The ability to obtain such data will be directly useful in both pure science and engineering applications. For example, hydrophobicity data could be used to refine the prediction of in-vitro protein aggregation. Protein surface hydrophobicity has been shown to be key to the mechanism of aggregation - a major obstacle in biochemical processing. The data could augment previous correlations of aggregation and the protein B22 - an osmotic second viral coefficient and measure of protein self interaction propensity.
The main challenges of this project are of an engineering nature: to develop a reliable and repeatable method using existing technology and equipment.
1. Hedberg, S. H. M., Heng, J. Y. Y., Williams, D. R., & Liddell, J. M. (2016). Micro scale self-interaction chromatography of proteins: A mAb case-study. Journal of Chromatography A, 1434, 57-63.
2. Tessier, P. M., Lenhoff, A. M., & Sandler, S. I. (2002). Rapid measurement of protein osmotic second virial coefficients by self-interaction chromatography. Biophysical journal, 82(3), 1620-1631.