PMID- 36068014
OWN - NLM
STAT- MEDLINE
DCOM- 20221010
LR  - 20221108
IS  - 1535-3907 (Electronic)
IS  - 1535-3893 (Print)
IS  - 1535-3893 (Linking)
VI  - 21
IP  - 10
DP  - 2022 Oct 7
TI  - Deep Single-Shot NanoLC-MS Proteome Profiling with a 1500 Bar UHPLC System, Long 
      Fully Porous Columns, and HRAM MS.
PG  - 2545-2551
LID - 10.1021/acs.jproteome.2c00270 [doi]
AB  - This study demonstrates how the latest ultrahigh-performance liquid 
      chromatography (UHPLC) technology can be combined with high-resolution 
      accurate-mass (HRAM) mass spectrometry (MS) and long columns packed with fully 
      porous particles to improve bottom-up proteomics analysis with nanoflow liquid 
      chromatography-mass spectrometry (nanoLC-MS) methods. The increased back 
      pressures from the UHPLC system enabled the use of 75 mum I.D. x 75 cm columns 
      packed with 2 mum particles at a typical 300 nL/min flow rate as well as elevated 
      and reduced flow rates. The constant pressure pump operation at 1500 bar reduced 
      sample loading and column washing/equilibration stages and overall overhead time, 
      which maximizes MS utilization time. The versatility of flow rate optimization to 
      balance the sensitivity, throughput with sample loading amount, and capability of 
      using longer gradients contributes to a greater number of peptide and protein 
      identifications for single-shot bottom-up proteomics experiments. The routine 
      proteome profiling and precise quantification of >7000 proteins with 
      single-shot nanoLC-MS analysis open possibilities for large-scale discovery 
      studies with a deep dive into the protein level alterations. Data are available 
      via ProteomeXchange with identifier PXD035665.
FAU - Zheng, Runsheng
AU  - Zheng R
AUID- ORCID: 0000-0002-5433-2540
AD  - Thermo Fisher Scientific, Dornierstrasse 4, 82110 Germering, Germany.
FAU - Stejskal, Karel
AU  - Stejskal K
AD  - IMP horizontal line Institute of Molecular Pathology, Campus-Vienna-Biocenter 1, A-1030 Vienna, 
      Austria.
AD  - IMBA horizontal line Institute of Molecular Biotechnology of the Austrian Academy of Sciences, 
      Dr. Bohr Gasse 3, A-1030 Vienna, Austria.
AD  - Gregor Mendel Institute of Molecular Plant Biology of the Austrian Academy of 
      Sciences, Dr. Bohr Gasse 3, A-1030 Vienna, Austria.
FAU - Pynn, Christopher
AU  - Pynn C
AD  - Thermo Fisher Scientific, Dornierstrasse 4, 82110 Germering, Germany.
FAU - Mechtler, Karl
AU  - Mechtler K
AUID- ORCID: 0000-0002-3392-9946
AD  - IMP horizontal line Institute of Molecular Pathology, Campus-Vienna-Biocenter 1, A-1030 Vienna, 
      Austria.
AD  - IMBA horizontal line Institute of Molecular Biotechnology of the Austrian Academy of Sciences, 
      Dr. Bohr Gasse 3, A-1030 Vienna, Austria.
AD  - Gregor Mendel Institute of Molecular Plant Biology of the Austrian Academy of 
      Sciences, Dr. Bohr Gasse 3, A-1030 Vienna, Austria.
FAU - Boychenko, Alexander
AU  - Boychenko A
AUID- ORCID: 0000-0003-1765-170X
AD  - Thermo Fisher Scientific, Dornierstrasse 4, 82110 Germering, Germany.
LA  - eng
GR  - I 3686/FWF_/Austrian Science Fund FWF/Austria
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20220906
PL  - United States
TA  - J Proteome Res
JT  - Journal of proteome research
JID - 101128775
RN  - 0 (Peptides)
RN  - 0 (Proteome)
SB  - IM
MH  - Chromatography, High Pressure Liquid/methods
MH  - Chromatography, Liquid
MH  - Mass Spectrometry
MH  - *Peptides/analysis
MH  - Porosity
MH  - *Proteome/analysis
PMC - PMC9552226
OTO - NOTNLM
OT  - Orbitrap
OT  - UHPLC
OT  - bottom-up proteomics
OT  - high-resolution accurate-mass
OT  - long nanocolumns
OT  - nanoLC-MS
OT  - single-shot analysis
COIS- The authors declare the following competing financial interest(s): Dr. Runsheng 
      Zheng, Dr. Christoper Pynn, Dr. Alexander Boychenko are employees of Thermo 
      Fisher Scientific. The other authors declare no competing financial interests.
EDAT- 2022/09/07 06:00
MHDA- 2022/10/12 06:00
PMCR- 2022/10/11
CRDT- 2022/09/06 20:02
PHST- 2022/09/07 06:00 [pubmed]
PHST- 2022/10/12 06:00 [medline]
PHST- 2022/09/06 20:02 [entrez]
PHST- 2022/10/11 00:00 [pmc-release]
AID - 10.1021/acs.jproteome.2c00270 [doi]
PST - ppublish
SO  - J Proteome Res. 2022 Oct 7;21(10):2545-2551. doi: 10.1021/acs.jproteome.2c00270. 
      Epub 2022 Sep 6.