Dataset Information

The characterization of column heating effect in nano-flow liquid chromatography mass spectrometry (nanoLC-MS)-based proteomics

ABSTRACT: In this study, we systematically compared the proteomics identification results obtained under room temperature and high temperature.

ORGANISM(S): Homo Sapiens

SUBMITTER: Minjia Tan

PROVIDER: PXD017016 | iProX | Thu Jan 09 00:00:00 GMT 2020

REPOSITORIES: iProX

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The characterization of column heating effect in nanoflow liquid chromatography mass spectrometry (nanoLC-MS)-based proteomics.

Li Bolin B Guo Fang F Hu Hao H Liu Ping P Tan Minjia M Pan Jianyi J Zhai Linhui L

Journal of mass spectrometry : JMS 20191216 1

Column heating strategy is often applied in nano-high-performance liquid chromatography-mass spectrometer (nanoHPLC-MS) platform for enhancing the analytical efficiency of peptides or proteins. Nonetheless, the influence effects of column heating in peptides or proteins identification still lack of deep understanding. In this study, a systematic comparison of room temperature (RT) and column heating of nanoHPLC was done. Based on the data, under column heating condition, the backpressure of nano ...[more]

PMID: 31840882

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Project description:Over the past 10 years, the bioenergy field has realized significant achievements that have encouraged many follow on efforts centered on biosynthetic production of fuel-like compounds. Key to the success of these efforts has been transformational developments in feedstock characterization and metabolic engineering of biofuel-producing microbes. Lagging far behind these advancements are analytical methods to characterize and quantify systems of interest to the bioenergy field. In particular, the utilization of proteomics, while valuable for identifying novel enzymes and diagnosing problems associated with biofuel-producing microbes, is limited by a lack of robustness and limited throughput. Nano-flow liquid chromatography coupled to high-mass accuracy, high-resolution mass spectrometers has become the dominant approach for the analysis of complex proteomic samples, yet such assays still require dedicated experts for data acquisition, analysis, and instrument upkeep. The recent adoption of standard flow chromatography (ca. 0.5 mL/min) for targeted proteomics has highlighted the robust nature and increased throughput of this approach for sample analysis. Consequently, we assessed the applicability of standard flow liquid chromatography for shotgun proteomics using samples from Escherichia coli and Arabidopsis thaliana, organisms commonly used as model systems for lignocellulosic biofuels research. Employing 120 min gradients with standard flow chromatography, we were able to routinely identify nearly 800 proteins from E. coli samples; while for samples from Arabidopsis, over 1,000 proteins could be reliably identified. An examination of identified peptides indicated that the method was suitable for reproducible applications in shotgun proteomics. Standard flow liquid chromatography for shotgun proteomics provides a robust approach for the analysis of complex samples. To the best of our knowledge, this study represents the first attempt to validate the standard flow approach for shotgun proteomics.

Dataset Information

The characterization of column heating effect in nano-flow liquid chromatography mass spectrometry (nanoLC-MS)-based proteomics

Publications

The characterization of column heating effect in nanoflow liquid chromatography mass spectrometry (nanoLC-MS)-based proteomics.

Similar Datasets

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