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

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• 作者：Jun Adachi ; Kazunari Hashiguchi ; Maiko Nagano ; Misako Sato ; Ayako Sato ; Kazuna Fukamizu ; Yasushi Ishihama ; Takeshi Tomonaga
• 刊名：Analytical Chemistry
• 出版年：2016
• 出版时间：August 16, 2016
• 年：2016
• 卷：88
• 期：16
• 页码：7899-7903
• 全文大小：337K
• 年卷期：0
• ISSN：1520-6882

文摘

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.