Open tubular lab-on-column/mass spectrometry for targeted proteomics of nanogram sample amounts.

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...

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Autores principales:	Hanne Kolsrud Hustoft, Tore Vehus, Ole Kristian Brandtzaeg, Stefan Krauss, Tyge Greibrokk, Steven Ray Wilson, Elsa Lundanes
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3a1fa308d0754cb4ba6ca682c549557f
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spelling	oai:doaj.org-article:3a1fa308d0754cb4ba6ca682c549557f2021-11-25T06:00:35ZOpen tubular lab-on-column/mass spectrometry for targeted proteomics of nanogram sample amounts.1932-620310.1371/journal.pone.0106881https://doaj.org/article/3a1fa308d0754cb4ba6ca682c549557f2014-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0106881https://doaj.org/toc/1932-6203A 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.Hanne Kolsrud HustoftTore VehusOle Kristian BrandtzaegStefan KraussTyge GreibrokkSteven Ray WilsonElsa LundanesPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 9, p e106881 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Hanne Kolsrud Hustoft Tore Vehus Ole Kristian Brandtzaeg Stefan Krauss Tyge Greibrokk Steven Ray Wilson Elsa Lundanes Open tubular lab-on-column/mass spectrometry for targeted proteomics of nanogram sample amounts.
description	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.
format	article
author	Hanne Kolsrud Hustoft Tore Vehus Ole Kristian Brandtzaeg Stefan Krauss Tyge Greibrokk Steven Ray Wilson Elsa Lundanes
author_facet	Hanne Kolsrud Hustoft Tore Vehus Ole Kristian Brandtzaeg Stefan Krauss Tyge Greibrokk Steven Ray Wilson Elsa Lundanes
author_sort	Hanne Kolsrud Hustoft
title	Open tubular lab-on-column/mass spectrometry for targeted proteomics of nanogram sample amounts.
title_short	Open tubular lab-on-column/mass spectrometry for targeted proteomics of nanogram sample amounts.
title_full	Open tubular lab-on-column/mass spectrometry for targeted proteomics of nanogram sample amounts.
title_fullStr	Open tubular lab-on-column/mass spectrometry for targeted proteomics of nanogram sample amounts.
title_full_unstemmed	Open tubular lab-on-column/mass spectrometry for targeted proteomics of nanogram sample amounts.
title_sort	open tubular lab-on-column/mass spectrometry for targeted proteomics of nanogram sample amounts.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/3a1fa308d0754cb4ba6ca682c549557f
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Open tubular lab-on-column/mass spectrometry for targeted proteomics of nanogram sample amounts.

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