The Use of Size Exclusion Chromatography to Monitor Protein Self-Assembly

The Use of Size Exclusion Chromatography to Monitor Protein Self-Assembly

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 ph...

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Bibliographic Details
Main Authors: Alaa Adawy, Matthew R. Groves
Format: Article
Language:English
Published: MDPI AG 2017-10-01
Series:Crystals
Subjects:
size exclusion chromatography
static light scattering
protein aggregation
protein nucleation
Online Access:https://www.mdpi.com/2073-4352/7/11/331
Description
Summary: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.
ISSN:2073-4352

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