The One Hour Human Proteome
Lia R. Serrano, Trenton M. Peters-Clarke, Tabiwang N. Arrey, Eugen Damoc, Margaret Lea Robinson, Noah M. Lancaster, Evgenia Shishkova, C. Moss, Anna Pashkova, Pavel Sinitcyn, Dain R. Brademan, Scott T. Quarmby, Amelia C. Peterson, Martin Zeller, Daniel Hermanson, Hamish Stewart, Christian Hock, Alexander Makarov, Vlad Zabrouskov, Joshua J. Coon
Abstract
We describe deep analysis of the human proteome in less than one hour. We achieve this expedited proteome characterization by leveraging state-of-the-art sample preparation, chromatographic separations, data analysis tools, and by using the new Orbitrap Astral mass spectrometer equipped with a quadrupole mass filter, a high-field Orbitrap mass analyzer, and an asymmetric track lossless (Astral) mass analyzer. The system offers high MS/MS acquisition speed of 200 Hz and detects hundreds of peptide sequences per second within data independent- or data-dependent acquisition modes of operation. The fast-switching capabilities of the new quadrupole complement the sensitivity and fast ion scanning of the Astral analyzer to enable narrow-bin data-independent analysis (DIA) methods. Over a 30-minute active chromatographic method consuming a total analysis time of 56 minutes, the Q-Orbitrap-Astral hybrid MS collects an average of 4,319 MS1 scans and 438,062 MS/MS scans per run, producing 235,916 peptide sequences (1% false discovery rate (FDR)). On average, each 30-minute analysis achieved detection of 10,411 protein groups (1% FDR). We conclude, with these results and alongside other recent reports, that the one-hour human proteome is within reach.