Probing Protein Denaturation during Size-Exclusion Chromatography Using Native Mass Spectrometry.

ABSTRACT: Size-exclusion chromatography employing aqueous mobile phases with volatile salts at neutral pH combined with electrospray-ionization mass spectrometry (SEC-ESI-MS) is a useful tool to study proteins in their native state. However, whether the applied eluent conditions actually prevent protein-stationary phase interactions, and/or protein denaturation, often is not assessed. In this study, the effects of volatile mobile phase additives on SEC retention and ESI of proteins were thoroughly investigated. Myoglobin was used as the main model protein, and eluents of varying ionic strength and pH were applied. The degree of interaction between protein and stationary phase was evaluated by calculating the SEC distribution coefficient. Protein-ion charge state distributions obtained during offline

SUBMITTER: Ventouri IK 

PROVIDER: S-EPMC7081181 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature