Tueshaus, J.; Eckert, S.; Fraefel, M.; Zhou, Y.; Pfeiffer, P.; Halves, C.; Fusco, F.; Weigl, J.; Hönikl, L.; Butenschön, V.; Todorova, R.; Rauert-Wunderlich, H.; The, M.; Rosenwald, A.; Heinemann, V.; Holch, J. W.; Meyer, B.; Weichert, W.; Mogler, C.; Kuhn, P.-H.; Küster, B.

Proteome profiling of formalin-fixed paraffin-embedded (FFPE) specimens has gained traction for the analysis of cancer tissue for the discovery of molecular biomarkers. However, reports so far focused on single cancer entities, comprised relatively few cases and did not assess the long-term performance of experimental workflows. Here, we did so by analyzing 1,220 tumors from six cancer entities processed over the course of three years. Key findings include the need for a new normalization method ensuring equal and reproducible sample loading for LC-MS/MS analysis across cohorts, showing that tumors can, on average, be profiled to a depth of >4,000 proteins and discovering that current software fails to process such large data sets. We report the first comprehensive pan-cancer proteome expression resource for FFPE material comprising 11,000 proteins which is of immediate utility to the scientific community by way of a web resource. It enables a range of analysis including quantitative comparisons of proteins between patients or cohorts or the discovery of protein fingerprints representing the tissue of origin, or proteins enriched in certain cancer entities.