Dataset Information

Evosep One Enables Robust Quantitative Deep Proteome Coverage using Tandem Mass Tags while Significantly Reducing Machine Time

ABSTRACT: The balance between comprehensively analyzing the proteome and using valuable mass spectrometry time is a genuine challenge in the field of proteomics. Multidimensional fractionation strategies have significantly increased proteome coverage, but often at the cost of increased mass analysis time, despite advances in mass spectrometer acquisition rates. Recently, the Evosep One liquid chromatography system was shown to analyze peptide samples in a high throughput manner without sacrificing in depth proteomics coverage. We demonstrate incorporation of Evosep One technology into our multiplexing workflow for quantitative analysis of tandem mass tag (TMT)-labeled non-small cell lung carcinoma (NSCLC) patient-derived xenografts (PDXs). Using a 30 samples per day Evosep workflow, >12,000 proteins were identified in 48 hours of mass spectrometry time, which is comparable to the number of proteins identified by our conventional concatenated EASY-nLC workflow in 67.5 hours. Shorter Evosep gradient lengths reduced the number of protein identifications by 10%, while decreasing mass analysis time by 50%. Thus, our Evosep workflow enables quantitative analysis of multiplexed samples in less time without conceding depth of proteome coverage.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell, Cell Culture

DISEASE(S): Lung Cancer

SUBMITTER: Jonathan Krieger

LAB HEAD: Michal F. Moran

PROVIDER: PXD012520 | Pride | 2019-04-04

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	EVOSEP - all Runs Combined.pride.mgf.gz	Mgf
	EVOSEP-allRunsCombined.mzML	Mzml
	Jiefei-500ug-PDX-Fractionation-Test-F1.raw	Raw
	Jiefei-500ug-PDX-Fractionation-Test-F10.raw	Raw
	Jiefei-500ug-PDX-Fractionation-Test-F11.raw	Raw

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Publications

Evosep One Enables Robust Deep Proteome Coverage Using Tandem Mass Tags while Significantly Reducing Instrument Time.

Krieger Jonathan R JR Wybenga-Groot Leanne E LE Tong Jiefei J Bache Nicolai N Tsao Ming S MS Moran Michael F MF

Journal of proteome research 20190412 5

The balance between comprehensively analyzing the proteome and using valuable mass spectrometry time is a genuine challenge in the field of proteomics. Multidimensional fractionation strategies have significantly increased proteome coverage, but often at the cost of increased mass analysis time, despite advances in mass spectrometer acquisition rates. Recently, the Evosep One liquid chromatography system was shown to analyze peptide samples in a high-throughput manner without sacrificing in-dept ...[more]

PMID: 30938160

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