Pseudo Internal Standard Approach for Label-Free Quantitative Proteomics

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• 作者：Tsuyoshi Tabata ; Toshitaka Sato ; Junro Kuromitsu ; Yoshiya Oda
• 刊名：Analytical Chemistry
• 出版年：2007
• 出版时间：November 15, 2007
• 年：2007
• 卷：79
• 期：22
• 页码：8440 - 8445
• 全文大小：140K
• 年卷期：v.79,no.22(November 15, 2007)
• ISSN：1520-6882

文摘

Quantitative proteome analysis has become a versatile toolto understand biological functions. Although stable isotope labeling is the most reliable method for quantitativemass spectrometry, preparation of isotope-labeled compounds is time-consuming and expensive. Simple label-free approaches have been introduced, but intensity-basedquantitation without standards is not generally acceptedas reliable, especially for small molecules. We havedeveloped a novel label-free quantitative proteome analysis using pseudo internal standards (PISs). This idea wasderived from northern blotting analysis, in which housekeeping genes are used as standards to normalize andcompare target gene expression levels in different samples.In many proteomics studies, most proteins do not changetheir expression levels under different conditions, andtherefore, these proteins can be employed as pseudointernal standards. This new approach is simple and doesnot require additional standards or labeling reagents. ThePIS method represents a novel approach for mass spectrometry-based comprehensive quantitatitation and mayalso be applicable to quantitative metabolome analysis.