Automated method for enrichment of newly synthesized proteins based on AHA labeling

In this study, a novel high throughput method was successfully developed for the automated enrichment of newly synthesized proteins (NSPs) from complex environments, like cell culture media supplemented with fetal bovine serum (FBS). The proposed automated method allows for the robust and effective simultaneous enrichment of dozen of samples in a period of 3 hours. The enrichment method was comprehensively evaluated and validated, and was applied to the mass spectrometry (MS) analysis of the FBS-rich secretome of melanoma A375 cells treated with the immune cytokines interferon alpha (IFN-α) and gamma (IFN-γ).