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Cost-effective gene...
Cost-effective generation of precise label-free quantitative proteomes in high-throughput by microLC and data-independent acquisition
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- Vowinckel, Jakob (författare)
- Biognosys AG,University Of Cambridge
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- Zelezniak, Aleksej, 1984 (författare)
- University Of Cambridge,Chalmers tekniska högskola,Chalmers University of Technology,Science for Life Laboratory (SciLifeLab)
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- Bruderer, Roland (författare)
- Biognosys AG
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- Mülleder, Michael (författare)
- University Of Cambridge,The Francis Crick Institute
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- Reiter, Lukas (författare)
- Biognosys AG
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- Ralser, M. (författare)
- The Francis Crick Institute,University Of Cambridge
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(creator_code:org_t)
- 2018-03-12
- 2018
- Engelska.
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Ingår i: Scientific Reports. - : Springer Science and Business Media LLC. - 2045-2322 .- 2045-2322. ; 8:1
- Relaterad länk:
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https://research.cha... (primary) (free)
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https://www.nature.c...
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https://doi.org/10.1...
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https://research.cha...
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https://research.cha...
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Abstract
Ämnesord
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- 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-A nd 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.
Ämnesord
- NATURVETENSKAP -- Kemi -- Analytisk kemi (hsv//swe)
- NATURAL SCIENCES -- Chemical Sciences -- Analytical Chemistry (hsv//eng)
- NATURVETENSKAP -- Data- och informationsvetenskap -- Bioinformatik (hsv//swe)
- NATURAL SCIENCES -- Computer and Information Sciences -- Bioinformatics (hsv//eng)
- TEKNIK OCH TEKNOLOGIER -- Elektroteknik och elektronik -- Datorsystem (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Electrical Engineering, Electronic Engineering, Information Engineering -- Computer Systems (hsv//eng)
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- ref (ämneskategori)
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