An Optimized Shotgun Strategy for the Rapid Generation of Comprehensive Human Proteomes
University of Copenhagen · Novo Nordisk Foundation · +1 more institution
Abstract
This study investigates the challenge of comprehensively cataloging the complete human proteome from a single-cell type using mass spectrometry (MS)-based shotgun proteomics. We modify a classical two-dimensional high-resolution reversed-phase peptide fractionation scheme and optimize a protocol that provides sufficient peak capacity to saturate the sequencing speed of modern MS instruments. This strategy enables the deepest proteome of a human single-cell type to date, with the HeLa proteome sequenced to a depth of ∼584,000 unique peptide sequences and ∼14,200 protein isoforms (∼12,200 protein-coding genes). This depth is comparable with next-generation RNA sequencing and enables the identification of…
Citation impact
- FWCI
- 32.21
- Percentile
- 100%
- References
- 72
Authors
12- DBDorte B. Bekker‐JensenCorresponding
University of Copenhagen, Novo Nordisk Foundation
- CDChristian D. Kelstrup
University of Copenhagen, Novo Nordisk Foundation
- TSTanveer S. Batth
University of Copenhagen, Novo Nordisk Foundation
- SCSara C. Buch-Larsen
University of Copenhagen, Novo Nordisk Foundation
- CHChrista Haldrup
Aarhus University Hospital
Topics & keywords
- Shotgun
- Shotgun proteomics
- Proteome
- Computational biology
- Computer science
- Biology
- Bioinformatics
- Proteomics