| Literature DB >> 17267305 |
Gustav Sundqvist1, Maria Stenvall, Helena Berglund, Jenny Ottosson, Harry Brumer.
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 disulfide bond formation.Entities:
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Year: 2007 PMID： 17267305 DOI： 10.1016/j.jchromb.2007.01.011
Source DB: PubMed Journal: J Chromatogr B Analyt Technol Biomed Life Sci ISSN： 1570-0232 Impact factor: 3.205