Dataset Information

Robust micro-flow LC-MS/MS for proteome analysis – 38,000 runs and counting

ABSTRACT: Micro-flow liquid chromatography tandem mass spectrometry (µLC–MS/MS) is increasingly becoming an alternative to nano-LC–MS/MS for the analysis of proteomes. We have recently demonstrated the potential of a µLC–MS/MS system operating with a 1 mm i.d. × 150 mm column and at a flow rate of 50 µl/min for high-throughput applications. Based on the analysis of ~38,000 samples measured on two instruments over the past two years, we show that the approach is extremely robust. Up to 1,500 analysis were performed within one month and >14,000 samples could be analyzed on a single column without loss of chromatographic performance. Samples included proteomes of cell lines, animal tissue and human body fluids, which were analyzed either with or without prior peptide fractionation or stable isotope labeling. We show that the µLC–MS/MS system is capable of measuring ~2,700 proteins from human plasma and ~5,200 proteins from human urine within one day, demonstrating its potential for in-depth as well as high-throughput clinical application.

INSTRUMENT(S): Q Exactive HF-X

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Urine, Blood Plasma, Epithelial Cell, Cell Culture

SUBMITTER: Yangyang Bian

LAB HEAD: Bernhard Kuster

PROVIDER: PXD023650 | Pride | 2021-02-22

REPOSITORIES: Pride

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Publications

Robust Microflow LC-MS/MS for Proteome Analysis: 38 000 Runs and Counting.

Bian Yangyang Y Bayer Florian P FP Chang Yun-Chien YC Meng Chen C Hoefer Stefanie S Deng Nan N Zheng Runsheng R Boychenko Oleksandr O Kuster Bernhard B

Analytical chemistry 20210217 8

Microflow liquid chromatography tandem mass spectrometry (μLC-MS/MS) is becoming a viable alternative to nanoflow LC-MS/MS for the analysis of proteomes. We have recently demonstrated the potential of such a system operating with a 1 mm i.d. × 150 mm column and at a flow rate of 50 μL/min for high-throughput applications. On the basis of the analysis of ∼38 000 samples measured on two instruments over the past two years, we now show that the approach is extremely robust. Up to 1500 analyses were ...[more]

PMID: 33596053

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Project description:Investigation of gene expression in cultured human skin epithelial keratinocytes (HaCaT) following non-thermal plasma treatment for 60 s, compared to untreated control. Non-thermal atmospheric pressure plasma has recently gained attention in the field of biomedical and clinical applications. In the area of plasma medicine research one promising approach is to promote wound healing by stimulation of cells involved. To understand basic molecular and cellular mechanisms triggered by plasma treatment we investigated biological effects of an argon plasma jet (kinpen) on human epithelial skin cells. Consequently, whole-genome microarrays were used to analyze this interaction in detail and identified a statistically significant modification of 3,274 genes including 1,828 up- and 1,446 down-regulated genes. Particularly, plasma-treated cells are characterized by differential expression of a considerable number of genes involved in the response to stress. In this regard, we found a plasma-dependent regulation of oxidative stress answer and increased expression of enzymes of the antioxidative defense system (e.g. 91 oxidoreductases). Our results demonstrate that plasma induces cell reactions of stress-sensing but also of proliferative nature. Consistent with gene expression changes as well as Ingenuity Pathway Analysis prediction, we propose that stimulating doses of plasma may protect epithelial skin cells in wound healing by promoting proliferation and differentiation. In conclusion, gene expression profiling may become an important tool in identifying plasma-related changes of gene expression. Our results underline the enormous clinical potential of plasma as a biomedical tool for stimulation of epithelial skin cells We investigated biological effects of an argon plasma jet on HaCaTs. Microarray were used to analyzed this interaction in detail. The transcripts analyzed in this study are further described in Schmidt et al. (2013): Non-thermal plasma treatment is associated with changes in transcriptome of human epithelial skin cells. Accepted in Journal Free Radical Research A study using total RNA recovered from at least 8 non-thermal plasma treated samples (300 ms*M-BM-5l/cell) and untreated HaCaT controls.

Dataset Information

Robust micro-flow LC-MS/MS for proteome analysis – 38,000 runs and counting

Publications

Robust Microflow LC-MS/MS for Proteome Analysis: 38 000 Runs and Counting.

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Robust Microflow LC-MS/MS for Proteome Analysis: 38 000 Runs and Counting.