Biomedical Engineering Reference
In-Depth Information
Chapter 2
Overview of Quantitative LC-MS Techniques for Proteomics
and Activitomics
John F. Timms and Pedro R. Cutillas
Abstract
LC-MS is a useful technique for protein and peptide quantification. In addition, as a powerful tool
for systems biology research, LC-MS can also be used to quantify post-translational modifications and
metabolites that reflect biochemical pathway activity. This review discusses the different analytical tech-
niques that use LC-MS for the quantification of proteins, their modifications and activities in a multiplex
manner.
Key words:
Mass spectrometry, proteomics, activitomics, quantification, isotope labeling, label free.
1.
Introduction
LC-MS has become a powerful molecular biology tool and
multiple strategies for peptide, protein and enzyme activity quan-
titation by LC-MS have been developed and applied to address
a wide range of biological questions.
Chapter 1
discusses the
concepts of biochemical pathway analysis using LC-MS. In this
chapter, we describe how to approach such types of analysis using
the different quantitative strategies available to the researcher.
LC-MS protein quantification strategies fall into two broad
categories: those that are label free and those which involve
protein/peptide labelling (known as differential mass tagging or
isotopic labelling). These strategies are mostly used for relative
quantification of proteins/peptides between two or more samples
of interest. However, these methods have also been adapted for
absolute quantitation, where the introduction of a known amount
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