Super-SILAC - A Precise and Easy to Use Met hod for Quantitative Proteome Analysis
Mass spectrometry (MS) has made great technical progress over the last years. As a consequence, it is increasingly applied to diverse biological problems, addressing questions in basic science as well as in clinical applications.
Accurate quantification of tumor proteomes is still a big challenge. Stable Isotope Labeling with Amino acids in Cell culture (SILAC) is currently the most accurate method and a very robust MS -based technology. Therefore, it is a valuable tool for quantitative proteomics. However, until now SILAC was not suitable for applications to human tissue samples since it requires the complete metabolic labeling of the entire proteome, which limits its use to cell cultures.
Prof. Matthias Mann and his coworkers at the Max Planck Institute for Biochemistry now managed to overcome these limitations. They developed a mix of multiple heavy SILAC labeled cells that accurately represents the tissue of interest (Super - SILAC). This mixture is then applied to the samples of interest as internal standard, enabling the quantification of essentially all peptides in the sample.
By using a mixture of labeled cells r ather than a single cell line, the quantification accuracy could be doubled. Because of the low preparation costs and its easy handling additionally to its accuracy, precision and reproducibility, Super - SILAC is a powerful tool for a wide range of applications: e.g. it is optimally suited for classifying tumors for personalized treatments, for discovery of new tumor markers or for proteome-related basic research studies.
PCT application filed: WO 2011042467 (A1)