biostudies-literatureKulak NANovo Nordisk UK Research FoundationNovo Nordisk Foundation Center for Protein ResearchNovo Nordisk FondenMax-Planck-Gesellschaft694-705https://www.ebi.ac.uk/biostudies/studies/S-EPMC5383787biostudies-literatureUnknown16(4)Recent advances in mass spectrometry (MS)-based proteomics now allow very deep coverage of cellular proteomes. To achieve near-comprehensive identification and quantification, the combination of a first HPLC-based peptide fractionation orthogonal to the on-line LC-MS/MS step has proven to be particularly powerful. This first dimension is typically performed with milliliter/min flow and relatively large column inner diameters, which allow efficient pre-fractionation but typically require peptide amounts in the milligram range. Here, we describe a novel approach termed "spider fractionator" in which the post-column flow of a nanobore chromatography system enters an eight-port flow-selector rotor valve. The valve switches the flow into different flow channels at constant time intervals, such Molecular & cellular proteomics : MCPLoss-less Nano-fractionator for High Sensitivity, High Coverage Proteomics.PMC5383787NNF15SA0018240PI Matthias MannNNF15CC0001Kulak NAGeyer PEMann MfalseLoss-less Nano-fractionator for High Sensitivity, High Coverage Proteomics.Recent advances in mass spectrometry (MS)-based proteomics now allow very deep coverage of cellular proteomes. To achieve near-comprehensive identification and quantification, the combination of a first HPLC-based peptide fractionation orthogonal to the on-line LC-MS/MS step has proven to be particularly powerful. This first dimension is typically performed with milliliter/min flow and relatively large column inner diameters, which allow efficient pre-fractionation but typically require peptide amounts in the milligram range. Here, we describe a novel approach termed "spider fractionator" in which the post-column flow of a nanobore chromatography system enters an eight-port flow-selector rotor valve. The valve switches the flow into different flow channels at constant time intervals, such 2017-01-01T00:00:00Z2017 Apr2026-05-30T01:28:42.429Z2019-03-27T02:40:40ZS-EPMC53837872812690010.1074/mcp.O116.065136