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  A Novel LC System Embeds Analytes in Pre-formed Gradients for Rapid, Ultra-robust Proteomics

Bache, N., Geyer, P. E., Bekker-Jensen, D. B., Hoerning, O., Falkenby, L., Treit, P. V., et al. (2018). A Novel LC System Embeds Analytes in Pre-formed Gradients for Rapid, Ultra-robust Proteomics. Molecular and Cellular Proteomics, 17(11): UNSP TIR118.000853, pp. 2284-2296. doi:10.1074/mcp.TIR118.000853.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0002-EE27-7 Version Permalink: http://hdl.handle.net/21.11116/0000-0002-EE28-6
Genre: Journal Article

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© 2018 Bache et al.

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 Creators:
Bache, Nicolai1, Author
Geyer, Philipp E.2, Author              
Bekker-Jensen, Dorte B.1, Author
Hoerning, Ole1, Author
Falkenby, Lasse1, Author
Treit, Peter V.2, Author              
Doll, Sophia2, Author              
Paron, Igor2, Author              
Müller, Johannes B.2, Author              
Meier, Florian2, Author              
Olsen, Jesper V.1, Author
Vorm, Ole1, Author
Mann, Matthias2, Author              
Affiliations:
1external, ou_persistent22              
2Mann, Matthias / Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565159              

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Free keywords: GO EXTRACTION TIPS; BIOMARKER DISCOVERY; MASS; PROTEIN; MS/MS; QUANTIFICATION; INTERACTOMES; WORKFLOW; DEPTH; STOPBiochemistry & Molecular Biology;
 Abstract: To further integrate mass spectrometry (MS)-based proteomics into biomedical research and especially into clinical settings, high throughput and robustness are essential requirements. They are largely met in high-flow rate chromatographic systems for small molecules but these are not sufficiently sensitive for proteomics applications. Here we describe a new concept that delivers on these requirements while maintaining the sensitivity of current nano-flow LC systems. Low-pressure pumps elute the sample from a disposable trap column, simultaneously forming a chromatographic gradient that is stored in a long storage loop. An auxiliary gradient creates an offset, ensuring the re-focusing of the peptides before the separation on the analytical column by a single high-pressure pump. This simplified design enables robust operation over thousands of sample injections. Furthermore, the steps between injections are performed in parallel, reducing overhead time to a few minutes and allowing analysis of more than 200 samples per day. From fractionated HeLa cell lysates, deep proteomes covering more than 130,000 sequence unique peptides and close to 10,000 proteins were rapidly acquired. Using this data as a library, we demonstrate quantitation of 5200 proteins in only 21 min. Thus, the new system - termed Evosep One analyzes samples in an extremely robust and high throughput manner, without sacrificing in depth proteomics coverage.

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Language(s): eng - English
 Dates: 2018
 Publication Status: Published online
 Pages: 13
 Publishing info: -
 Table of Contents: -
 Rev. Method: -
 Identifiers: ISI: 000451799900015
DOI: 10.1074/mcp.TIR118.000853
 Degree: -

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Project name : MSmed project
Grant ID : 686547
Funding program : Horizon 2020 (H2020)
Funding organization : European Commission (EC)

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Title: Molecular and Cellular Proteomics
Source Genre: Journal
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Publ. Info: Bethesda, MD : American Society for Biochemistry and Molecular Biology
Pages: - Volume / Issue: 17 (11) Sequence Number: UNSP TIR118.000853 Start / End Page: 2284 - 2296 Identifier: ISSN: 1535-9476
CoNE: https://pure.mpg.de/cone/journals/resource/111035577487002