MS04-P11 Size exclusion chromatography as a lab-based indicative for protein self-assembly prior to nucleation Alaa Adawy ( Laboratory of High-Resolution Transmission Electron Microscopy, Scientific-Technical Services, University of Oviedo-CINN, Oviedo, Spain) Matthew Groves (Department of Drug Design, Structural Biology Unit, Groningen Research Institute of Pharmacy, University of Groningen, Groningen, Netherlands (Holland, Europe))email: hassanalaa@uniovi.esThe potential use of size exclusion chromatography (SEC) as a lab-based scanning methodology to monitor protein self-assembly prior to nucleation is the subject of this study. The effect of the mobile phase ionic strength and protein concentration on the output of SEC experiments was exploerd using high-resolution size exclusion chromatography (SEC) coupled with static light scattering (SLS) analyses. SLS is capable of monitoring abrupt changes in the molar masses and the results highlight the effect of small changes in the mobile phase composition on the estimation of molar masses calculated from retention time-based calibration curve compared with those obtained from SLS analysis. By comparing the SLS data with the SEC chromatograms, we show that SEC can provide helpful information on the protein aggregation state as macromolecules approach known precipitation points in their phase diagrams. This study implies that SEC can be usefully used to study phase diagrams of different proteins.  References:

Adawy, A & Groves, M.R. (2017) Crystals 7(11), 331; doi:10.3390/cryst7110331
Keywords: size exclusion chromatography, static light scattering, protein nucleation