dc.date.accessioned | 2015-04-10T13:41:37Z | |
dc.date.available | 2015-04-10T13:41:37Z | |
dc.date.created | 2014-12-03T16:32:16Z | |
dc.date.issued | 2014 | |
dc.identifier.citation | Hustoft, Hanne Kolsrud Vehus, Tore Brandtzæg, Ole Kristian Merkesvik Krauss, Stefan Greibrokk, Tyge Wilson, Steven Ray Haakon Lundanes, Elsa . Open tubular lab-on-column/mass spectrometry for targeted proteomics of nanogram sample amounts. PLoS ONE. 2014, 9(9) | |
dc.identifier.uri | http://hdl.handle.net/10852/43547 | |
dc.description.abstract | A novel open tubular nanoproteomic platform featuring accelerated on-line protein digestion and high-resolution nano liquid chromatography mass spectrometry (LC-MS) has been developed. The platform features very narrow open tubular columns, and is hence particularly suited for limited sample amounts. For enzymatic digestion of proteins, samples are passed through a 20 µm inner diameter (ID) trypsin + endoproteinase Lys-C immobilized open tubular enzyme reactor (OTER). Resulting peptides are subsequently trapped on a monolithic pre-column and transferred on-line to a 10 µm ID porous layer open tubular (PLOT) liquid chromatography LC separation column. Wnt/ß-catenein signaling pathway (Wnt-pathway) proteins of potentially diagnostic value were digested+detected in targeted-MS/MS mode in small cell samples and tumor tissues within 120 minutes. For example, a potential biomarker Axin1 was identifiable in just 10 ng of sample (protein extract of ~1,000 HCT15 colon cancer cells). In comprehensive mode, the current OTER-PLOT set-up could be used to identify approximately 1500 proteins in HCT15 cells using a relatively short digestion+detection cycle (240 minutes), outperforming previously reported on-line digestion/separation systems. The platform is fully automated utilizing common commercial instrumentation and parts, while the reactor and columns are simple to produce and have low carry-over. These initial results point to automated solutions for fast and very sensitive MS based proteomics, especially for samples of limited size. | en_US |
dc.language | EN | |
dc.language.iso | en | en_US |
dc.publisher | Public Library of Science (PLoS) | |
dc.rights | Attribution 4.0 International | |
dc.rights.uri | http://creativecommons.org/licenses/by/4.0/ | |
dc.title | Open tubular lab-on-column/mass spectrometry for targeted proteomics of nanogram sample amounts | en_US |
dc.type | Journal article | en_US |
dc.creator.author | Hustoft, Hanne Kolsrud | |
dc.creator.author | Vehus, Tore | |
dc.creator.author | Brandtzæg, Ole Kristian Merkesvik | |
dc.creator.author | Krauss, Stefan | |
dc.creator.author | Greibrokk, Tyge | |
dc.creator.author | Wilson, Steven Ray Haakon | |
dc.creator.author | Lundanes, Elsa | |
cristin.unitcode | 185,15,12,0 | |
cristin.unitname | Kjemisk institutt | |
cristin.ispublished | true | |
cristin.fulltext | original | |
cristin.qualitycode | 1 | |
dc.identifier.cristin | 1180644 | |
dc.identifier.bibliographiccitation | info:ofi/fmt:kev:mtx:ctx&ctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.jtitle=PLoS ONE&rft.volume=9&rft.spage=&rft.date=2014 | |
dc.identifier.jtitle | PLoS ONE | |
dc.identifier.volume | 9 | |
dc.identifier.issue | 9 | |
dc.identifier.doi | http://dx.doi.org/10.1371/journal.pone.0106881 | |
dc.identifier.urn | URN:NBN:no-47910 | |
dc.type.document | Tidsskriftartikkel | en_US |
dc.type.peerreviewed | Peer reviewed | |
dc.source.issn | 1932-6203 | |
dc.identifier.fulltext | Fulltext https://www.duo.uio.no/bitstream/handle/10852/43547/2/journal-pone-0106881vehus.pdf | |
dc.type.version | PublishedVersion | |
cristin.articleid | e106881 | |