The use of size exclusion chromatography to monitor protein self-assembly
Adawy, Alaa; Groves, Matthew R.;
Abstract
High resolution size exclusion chromatography (SEC) coupled with static light scattering (SLS) analyses were conducted to study the effect of the mobile phase ionic strength and protein concentration on the output of SEC experiments. The results highlight the effect of small changes in the mobile phase composition on the estimation of molar masses estimated 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 suggests the potential use of SEC as an easily accessible lab-based scanning methodology to monitor protein self-assembly prior to nucleation and crystallization. Implications for the use of SEC to study protein phase diagrams are discussed.
Other data
Title | The use of size exclusion chromatography to monitor protein self-assembly | Authors | Adawy, Alaa ; Groves, Matthew R. | Keywords | Protein aggregation | Protein nucleation | Size exclusion chromatography | Static light scattering | Issue Date | 1-Nov-2017 | Publisher | MDPI AG | Journal | Crystals | Volume | 7 | Issue | 11 | ISSN | 2073-4352 | DOI | 10.3390/cryst7110331 | Scopus ID | 2-s2.0-85033595609 | Web of science ID | WOS:000416601500010 |
Recommend this item
Similar Items from Core Recommender Database
Items in Ain Shams Scholar are protected by copyright, with all rights reserved, unless otherwise indicated.