A streamlined platform for high-content functional proteomics of primary human specimens

摘要: Achieving information content of satisfactory breadth and depth remains a formidable challenge for proteomics. This problem is particularly relevant to the study primary human specimens, such as tumor biopsies, which are heterogeneous finite quantity. Here we present functional proteomics strategy that unites activity-based protein profiling multidimensional identification technologies (ABPP-MudPIT) streamlined analysis samples. convergent platform involves rapid initial phase, in enzyme activity signatures generated classification samples, followed by in-depth representative members from each class. Using this two-tiered approach, identified more than 50 activities breast tumors, nearly third represent previously uncharacterized proteins. Comparison with cDNA microarrays revealed enzymes whose activity, but not mRNA expression, depicted class, underscoring power ABPP-MudPIT discovery new markers disease may evade detection other molecular methods.