Improved Proteome and Phosphoproteome Analysis on a Cation Exchanger by a Combined Acid and Salt Gradient

Currently used elution methods for strong cation exchange (SCX) chromatography are based on two principles: salt and pH gradient. In this paper, we report the first observation of peptide elution by acid gradient. The degree of peptide separation using C18-SCX StageTip was greatly improved by our ac...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2016-08, Vol.88 (16), p.7899-7903
Main Authors: Adachi, Jun, Hashiguchi, Kazunari, Nagano, Maiko, Sato, Misako, Sato, Ayako, Fukamizu, Kazuna, Ishihama, Yasushi, Tomonaga, Takeshi
Format: Article
Language:English
Subjects:
Analytical chemistry
Cation exchanging
Cations - chemistry
Chromatography
Chromatography, Ion Exchange
Elution
Exchange
HeLa Cells
Humans
Hydrogen-Ion Concentration
Peptides
Peptides - chemistry
Peptides - isolation & purification
Proteins
Proteome
Salt
Salts - chemistry
Separation
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Summary:Currently used elution methods for strong cation exchange (SCX) chromatography are based on two principles: salt and pH gradient. In this paper, we report the first observation of peptide elution by acid gradient. The degree of peptide separation using C18-SCX StageTip was greatly improved by our acid and salt-based elution method compared with a salt-based elution method. This development enabled us to identify over 22 000 phosphopeptides from 2 mg of protein without labor-intensive sample preparation. Our method is simple, robust, scalable, and low-cost and can be easily implemented without any special equipment or techniques.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.6b01232