Branca, Rui M MOrre, Lukas MJohansson, Henrik JGranholm, ViktorHuss, MikaelPerez-Bercoff, AsaForshed, JennyKäll, LukasLehtiö, Janne2015-12-071548-7091http://hdl.handle.net/1885/25698We present a liquid chromatography-mass spectrometry (LC-MS)-based method permitting unbiased (gene prediction-independent) genome-wide discovery of protein-coding loci in higher eukaryotes. Using high-resolution isoelectric focusing (HiRIEF) at the peptide level in the 3.7-5.0 pH range and accurate peptide isoelectric point (pI) prediction, we probed the six-reading-frame translation of the human and mouse genomes and identified 98 and 52 previously undiscovered protein-coding loci, respectively. The method also enabled deep proteome coverage, identifying 13,078 human and 10,637 mouse proteins.201410.1038/nmeth.27322015-12-07