title: The Q Exactive HF, a Benchtop mass spectrometer with a pre-filter, high-performance quadrupole and an ultra-high-field Orbitrap analyzer.
abstract: The quadrupole Orbitrap mass spectrometer (Q Exactive) made a powerful proteomics instrument available in a benchtop format. It significantly boosted the number of proteins analyzable per hour and has now evolved into a proteomics analysis workhorse for many laboratories. Here we describe the Q Exactive Plus and Q Exactive HF mass spectrometers, which feature several innovations in comparison to the original Q Exactive instrument. A low-resolution pre-filter has been implemented within the injection flatapole, preventing unwanted ions from entering deep into the system, and thereby increasing its robustness. A new segmented quadrupole, with higher fidelity of isolation efficiency over a wide range of isolation windows, provides an almost 2-fold improvement of transmission at narrow isolation widths. Additionally, the Q Exactive HF has a compact Orbitrap analyzer, leading to higher field strength and almost doubling the resolution at the same transient times. With its very fast isolation and fragmentation capabilities, the instrument achieves overall cycle times of 1 s for a top 15 to 20 higher energy collisional dissociation method. We demonstrate the identification of 5000 proteins in standard 90-min gradients of tryptic digests of mammalian cell lysate, an increase of over 40% for detected peptides and over 20% for detected proteins. Additionally, we tested the instrument on peptide phosphorylation enriched samples, for which an improvement of up to 60% class I sites was observed.
authors.affiliation: From the ‡Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Am Klopferspitz 18, D-82152 Martinsried, Germany;
authors.forename: Richard Alexander
authors.initials: RA
authors.lastname: Scheltema
authors.affiliation: §Thermo Fisher Scientific (Bremen) GmbH, Hanna-Kunath-Strasse 11, 28199 Bremen, Germany.
authors.forename: Jan-Peter
authors.initials: JP
authors.lastname: Hauschild
authors.affiliation: §Thermo Fisher Scientific (Bremen) GmbH, Hanna-Kunath-Strasse 11, 28199 Bremen, Germany.
authors.forename: Oliver
authors.initials: O
authors.lastname: Lange
authors.affiliation: From the ‡Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Am Klopferspitz 18, D-82152 Martinsried, Germany;
authors.forename: Daniel
authors.initials: D
authors.lastname: Hornburg
authors.affiliation: §Thermo Fisher Scientific (Bremen) GmbH, Hanna-Kunath-Strasse 11, 28199 Bremen, Germany.
authors.forename: Eduard
authors.initials: E
authors.lastname: Denisov
authors.affiliation: §Thermo Fisher Scientific (Bremen) GmbH, Hanna-Kunath-Strasse 11, 28199 Bremen, Germany.
authors.forename: Eugen
authors.initials: E
authors.lastname: Damoc
authors.affiliation: §Thermo Fisher Scientific (Bremen) GmbH, Hanna-Kunath-Strasse 11, 28199 Bremen, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Kuehn
authors.affiliation: §Thermo Fisher Scientific (Bremen) GmbH, Hanna-Kunath-Strasse 11, 28199 Bremen, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Makarov
authors.affiliation: From the ‡Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Am Klopferspitz 18, D-82152 Martinsried, Germany; mmann@biochem.mpg.de.
authors.forename: Matthias
authors.initials: M
authors.lastname: Mann
mesh_terms: D000595:Amino Acid Sequence; D005374:Filtration; D017022:Flow Injection Analysis; D006367:HeLa Cells; D006801:Humans; D007477:Ions; D013058:Mass Spectrometry / Q000295:instrumentation* / Q000379:methods; D008969:Molecular Sequence Data; D010750:Phosphoproteins / Q000302:isolation & purification*; D010766:Phosphorylation; D012680:Sensitivity and Specificity; D013997:Time Factors; D014357:Trypsin / Q000737:chemistry
chemical_list: D007477:Ions; D010750:Phosphoproteins; D014357:Trypsin