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A general, robust method for the quality control of intact proteins using LC-ESI-MS

TitleA general, robust method for the quality control of intact proteins using LC-ESI-MS
Publication TypeJournal Article
Year of Publication2007
AuthorsSundqvist, G, Stenvall, M, Berglund, H, Ottosson, J, Brumer, H
JournalJOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
Volume852
Pagination188-194
Date PublishedJUN 1
Type of ArticleArticle
ISSN1570-0232
Abstract

A simple and robust method for the routine quality control of intact proteins based on liquid chromatography coupled to electrospray ionization mass spectrometry (LC-ESI-MS) is presented. A wide range of prokaryotic and eukaryotic proteins expressed recombinantly in Escherichia coli or Pichia pastoris has been analyzed with medium- to high-throughput with on-line desalting from multi-well sample plates. Particular advantages of the method include fast chromatography and short cycle times, the use of inexpensive trapping/desalting columns, low sample carryover, and the ability to analyze proteins with masses ranging from 5 to 100 kDa with greater than 50 ppm accuracy. Moreover, the method can be readily coupled with optimized chemical reduction and alkylation steps to facilitate the analysis of denatured or incorrectly folded proteins (e.g., recombinant proteins sequestered in E. coli inclusion bodies) bearing cysteine residues, which otherwise form intractable multimers and non-specific adducts by disultide bond formation. (C) 2007 Elsevier B.V.. All rights reserved.

DOI10.1016/j.jchromb.2007.01.011