Abstract

In the era of precision medicine, there is an increasing need to measure several thousand proteins expressed in minimal amount of fresh-frozen biopsy tissue samples from clinical cohorts. Here we present the detailed protocol for high-throughput proteomic analysis of less than 1 mg fresh-frozen tissue sample using pressure cycling technology (PCT), SWATH mass spectrometry, and OpenSWATH workflow. The PCT procedures enable simultaneous analysis of 16 biopsy tissue samples per batch, followed by SWATH analysis of about 15 samples per mass spectrometer per working day. The thus generated SWATH maps can be perpetually analyzed in silico, offering a practical solution for digital biobanking