Integrated enzyme reactor and high resolving chromatography in “sub-chip” dimensions for sensitive protein mass spectrometry View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2013-12

AUTHORS

Hanne Kolsrud Hustoft, Ole Kristian Brandtzaeg, Magnus Rogeberg, Dorna Misaghian, Silje Bøen Torsetnes, Tyge Greibrokk, Léon Reubsaet, Steven Ray Wilson, Elsa Lundanes

ABSTRACT

Reliable, sensitive and automatable analytical methodology is of great value in e.g. cancer diagnostics. In this context, an on-line system for enzymatic cleavage of proteins, subsequent peptide separation by liquid chromatography (LC) with mass spectrometric detection has been developed using "sub-chip" columns (10-20 μm inner diameter, ID). The system could detect attomole amounts of isolated cancer biomarker progastrin-releasing peptide (ProGRP), in a more automatable fashion compared to previous methods. The workflow combines protein digestion using an 20 μm ID immobilized trypsin reactor with a polymeric layer of 2-hydroxyethyl methacrylate-vinyl azlactone (HEMA-VDM), desalting on a polystyrene-divinylbenzene (PS-DVB) monolithic trap column, and subsequent separation of resulting peptides on a 10 μm ID (PS-DVB) porous layer open tubular (PLOT) column. The high resolution of the PLOT columns was maintained in the on-line system, resulting in narrow chromatographic peaks of 3-5 seconds. The trypsin reactors provided repeatable performance and were compatible with long-term storage. More... »

PAGES

3511

References to SciGraph publications

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/srep03511

DOI

http://dx.doi.org/10.1038/srep03511

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1027295928

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/24336509


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