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• W2805896072 abstract "Quantitative proteomics is key for basic research, but needs improvements to satisfy an increasing demand for large sample series in diagnostics, academia and industry. A switch from nanoflowrate to microflowrate chromatography can improve throughput and reduce costs. However, concerns about undersampling and coverage have so far hampered its broad application. We used a QTOF mass spectrometer of the penultimate generation (TripleTOF5600), converted a nanoLC system into a microflow platform, and adapted a SWATH regime for large sample series by implementing retention time- and batch correction strategies. From 3 µg to 5 µg of unfractionated tryptic digests that are obtained from proteomics-typical amounts of starting material, microLC-SWATH-MS quantifies up to 4000 human or 1750 yeast proteins in an hour or less. In the acquisition of 750 yeast proteomes, retention times varied between 2% and 5%, and quantified the typical peptide with 5-8% signal variation in replicates, and below 20% in samples acquired over a five-months period. Providing precise quantities without being dependent on the latest hardware, our study demonstrates that the combination of microflow chromatography and data-independent acquisition strategies has the potential to overcome current bottlenecks in academia and industry, enabling the cost-effective generation of precise quantitative proteomes in large scale." @default.
• W2805896072 created "2018-06-13" @default.
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• W2805896072 date "2018-03-12" @default.
• W2805896072 modified "2023-10-10" @default.
• W2805896072 title "Cost-effective generation of precise label-free quantitative proteomes in high-throughput by microLC and data-independent acquisition" @default.
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• W2805896072 doi "https://doi.org/10.1038/s41598-018-22610-4" @default.
• W2805896072 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/5847575" @default.
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• W2805896072 hasPublicationYear "2018" @default.
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