Stability and phase transitions of protein solutions.

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Project description from Risk Form for you to adapt: The stability of proteins is important for normal biological function and for the storage and delivery of biopharmaceutical drugs. However, assessing stability can be challenging, since it relates both to the structural stability of the protein and its solution stability. Much of the challenge relates to the relative sparsity of techniques that can adequately address both, generally leading to a significant analytical burden for the testing and characterisation of protein stability in the biopharmaceutical industry. However, some techniques such as intrinsic fluorescence and circular dichroism (CD) can offer significant advantages over other approaches, in that they are both informative are require relatively little sample. Indirectly, the position of phase transition of protein can provide information on their relative stability and could improve predictability of protein solution stability. Liquid-liquid phase separation (LLPS) of crystallin proteins was described in the eye lens over 30 years ago. LLPS is an important phase transition in biology, and is metastable with respect to crystallization, so is therefore also a good proxy for protein stability. LLPS occurs as a result of net short ranged attractive inter-protein interactions between the gamma crystallin proteins found in the eye lens and occurs in both purified human gamma D crystallin (HGD) solutions in-vitro and in the complex mixture of alpha, beta and gamma crystallins found in eye lens fibres. Recently, due to renewed interest in studying liquid phase separation in cellular organelles, these processes have received significant interest in the literature. In this project, which will be conducted in collaboration with Applied Photophysics, we will explore stability and phase transitions of several globular and synthetic proteins using a combination of techniques, including those being developed by Applied Photophysics. In using instruments under development at Applied Photophysics to explore protein phase transitions and unfolding, we will help to guide their product development and by publishing our work, provide users with examples of how these instruments can be employed in their own research. This project falls within the EPSRC Physical Science research area.