Characterizing Mitochondrial Proteomes in Primary Leukaemic Cells and Cell Lines

摘要

Proteomic studies of cancer cells have made considerable progress over the last few years with the development of more sensitive mass spectrometry instruments. The combination of these instruments with improved and innovative approaches in sample preparation, including sub-cellular fractionation and protein separation techniques, is now allowing the characterisation and analysis of organelles from primary cancer cells as well as cultured cells. The targeted proteomic analysis of purified mitochondria and other sub-cellular organelles can potentially provide the basis for the development of new diagnostic and therapeutic targets. The anti-TOM20 innovative approach to mitochondrial purification combines differential centrifugation, a well-established and robust technique, with the specificity of immuno-precipitation. Purifications can be performed within a day, without the need for costly high-speed centrifuges, and samples prepared for mass spectrometry analysis within hours. The technique is highly specific with very low amounts of contamination and can be adapted to different cell types.