title: Evaluation of the compact high-field orbitrap for top-down proteomics of human cells.
abstract: Mass spectrometry based proteomics generally seeks to identify and fully characterize protein species with high accuracy and throughput. Recent improvements in protein separation have greatly expanded the capacity of top-down proteomics (TDP) to identify a large number of intact proteins. To date, TDP has been most tightly associated with Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry. Here, we couple the improved separations to a Fourier-transform instrument based not on ICR but using the Orbitrap Elite mass analyzer. Application of this platform to H1299 human lung cancer cells resulted in the unambiguous identification of 690 unique proteins and over 2000 proteoforms identified from proteins with intact masses<50 kDa. This is an early demonstration of high throughput TDP (>500 identifications) in an Orbitrap mass spectrometer and exemplifies an accessible platform for whole protein mass spectrometry.
authors: Department of Chemistry, Northwestern University, Evanston, Illinois, United States.
authors: Dorothy R
authors: DR
authors: Ahlf
authors: Philip D
authors: PD
authors: Compton
authors: John C
authors: JC
authors: Tran
authors: Bryan P
authors: BP
authors: Early
authors: Paul M
authors: PM
authors: Thomas
authors: Neil L
authors: NL
authors: Kelleher
mesh_terms: D000595:Amino Acid Sequence; D002166:Camptothecin / Q000494:pharmacology; D059447:Cell Cycle Checkpoints; D045744:Cell Line, Tumor; D016922:Cellular Senescence; D004249:DNA Damage; D005583:Fourier Analysis; D006801:Humans; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D010449:Peptide Mapping; D020543:Proteome / Q000737:chemistry* / Q000302:isolation & purification; D040901:Proteomics; D012015:Reference Standards; D053719:Tandem Mass Spectrometry / Q000592:standards*; D059003:Topoisomerase Inhibitors / Q000494:pharmacology
chemical_list: D010446:Peptide Fragments; D020543:Proteome; D059003:Topoisomerase Inhibitors; D002166:Camptothecin