An iTRAQ-RPLC-MS/MS approach for protein differential expression profiling of MCF7 breast cancer cells: Towards biomarker discovery

摘要

Biomarker discovery and screening using novel proteomic technologies is an area that is attracting increased interest. Recently, mass spectrometry (MS) based methods have been implemented for quantitative proteomics. Two main strategies, namely, label-free and stable isotope labeling, have been established. Among the stable isotope labeling strategies, iTRAQ (isotope tags for relative and absolute quantitation), in particular, has received much attention. The method involves peptide labeling with isobaric tags at the N-terminus and the lysine side chains. Upon MS/MS fragmentation, signature ions are generated for quantitative purposes. With this technique, up to 4-8 different samples can be simultaneously quantified. Here, we report an iTRAQ-reversed phase liquid chromatography (RPLC)-MS/MS method for protein differential expression profiling of the MCF7 breast cancer cells.