2021
DOI: 10.1101/2021.11.28.470272
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Deep proteome profiling with reduced carry over using superficially porous microfabricated nanoLC columns

Abstract: In the field of LC-MS based proteomics, increases in sampling depth and proteome coverage have mainly been accomplished by rapid advances in mass spectrometer technology. The comprehensiveness and quality of data that can be generated do however also depend on the performance provided by nano liquid chromatography (nanoLC) separations. Proper selection of reversed-phase separation columns can be of paramount importance to provide the MS instrument with peptides at the highest possible concentration and separat… Show more

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Cited by 5 publications
(11 citation statements)
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“…To our surprise, a striking protein ID gap between an injection amount of 10 ng and 50 ng was observed, which had not been reported by earlier studies in our lab. 10 In addition, increasing the peptide load led to a less pronounced increase in protein IDs. Only for longer gradients, the protein ID gap between different peptide loads showed a wider spread.…”
Section: Resultsmentioning
confidence: 99%
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“…To our surprise, a striking protein ID gap between an injection amount of 10 ng and 50 ng was observed, which had not been reported by earlier studies in our lab. 10 In addition, increasing the peptide load led to a less pronounced increase in protein IDs. Only for longer gradients, the protein ID gap between different peptide loads showed a wider spread.…”
Section: Resultsmentioning
confidence: 99%
“…This enables fast loading, washing and conditioning of the column pre-and post-analysis increasing sample throughput. 10,11 This is combined with a wide window acquisition (WWA) DDA method, which merges the strengths of DDA and DIA. As indicated by the name, we use a broad isolation window of ≥4 m/z in an otherwise data dependent precursor selection.…”
Section: Introductionmentioning
confidence: 99%
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“…The recently reported proteomics applications of such columns demonstrated superb kinetics, which microfluidic columns cannot compare with at the current stage. 52,53 In practical terms, however, the limited loadability of the pillar array and open tubular columns cannot compete with particle-packed columns at this moment. In this sense, the microfluidic packed meter-long columns have a practical transferability to proteomics laboratories although more analytical time is needed to realize deep-mining proteomics.…”
Section: ■ Results and Discussionmentioning
confidence: 99%