Protein Sciences in Drug Discovery 2022
Poster
8

Detergent-free purification of membrane proteins using polymer lipid particle (PoLiPa) technology for use in Cryo-EM

Abstract

The structural and biophysical analysis of membrane proteins such as GPCRs, ion channels and transport proteins is essential for modern drug discovery but has been limited by access to high quality purified protein and limitations of crystal formation. Contemporary techniques that rely upon detergent solubilisation of membrane proteins are greatly hindered by the challenges, time and costs associated with the bespoke method development needed for each purification. Domainex are developing a platform approach to membrane protein purification and structural analysis, enabled by polymer-based strategies, such as styrene maleic acid (SMA), that alleviate these issues. These solubilise cellular membranes and encapsulate membrane proteins that continually remain in their original native lipid environment, forming polymer lipid particles (PoLiPas). Thus, the membrane protein is presented in its folded state in solution, allowing purification using conventional techniques. Here, we showcase our system for rapid and generic preparation of high quality PoLiPa-purified proteins. To take advantage of this native lipid environment for structural studies, we also show how Cryo-EM can be used as a complimentary technique to visualise PoLiPa discs.

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The European Laboratory Research & Innovation Group Our Vision : To provide outstanding, leading edge knowledge to the life sciences community on an open access basis

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