Enhancing cognate target elution efficiency in gel-free chemical proteomics

• Main Authors: Branka Radic-Sarikas, Uwe Rix, Alexey Stukalov, Manuela Gridling, André C. Müller, Jacques Colinge, Giulio Superti-Furga, Keiryn L. Bennett
• Format: Article
• Language: English
• Published: Elsevier 2015-12-01
• Series: EuPA Open Proteomics
• Online Access: http://www.sciencedirect.com/science/article/pii/S2212968515300210

Gel-free liquid chromatography mass spectrometry coupled to chemical proteomics is a powerful approach for characterizing cellular target profiles of small molecules. We have previously described a fast and efficient elution protocol; however, altered target profiles were observed. We hypothesised that elution conditions critically impact the effectiveness of disrupting drug-protein interactions. Thus, a number of elution conditions were systematically assessed with the aim of improving the recovery of all classes of proteins whilst maintaining compatibility with immunoblotting procedures. A double elution with formic acid combined with urea emerged as the most efficient and generically applicable elution method for chemical proteomics Keywords: Chemical proteomics, Mass spectrometry gel-free, Double elution, Dasatinib, Sunitinib