Pseudo-MRM method for the selective detection of truncated tissue factor

A pseudo-MRM method using high-resolution mass spectrometry is presented for the specific detection of truncated tissue factor (tTF), retargeted to tumor vasculature by GNGRAHA peptide (tTF-NGR), in serum. In order to increase sensitivity, serum proteins are pre-separated by 1D-gel electrophoresis a...

Authors: König, Simone
Harrach, Saliha
Berdel, Wolfgang E.
Schwöppe, Christian
Division/Institute:FB 05: Medizinische Fakultät
Document types:Article
Media types:Text
Publication date:2017
Date of publication on miami:02.11.2017
Modification date:02.11.2017
Source:Mercator Journal of Biomolecular Analysis, 1 (2017) 2, S. 22-32
Edition statement:[Electronic ed.]
Subjects:mass spectrometry; protein quantification; serum; tumor; LC/MS
DDC Subject:572: Biochemie
License:CC BY-SA 3.0 DE
Language:English
Format:PDF document
URN:urn:nbn:de:hbz:6-00269468510
Permalink:http://nbn-resolving.de/urn:nbn:de:hbz:6-00269468510
Digital documents:mercator-journal_2017_1_2_22-32.pdf

A pseudo-MRM method using high-resolution mass spectrometry is presented for the specific detection of truncated tissue factor (tTF), retargeted to tumor vasculature by GNGRAHA peptide (tTF-NGR), in serum. In order to increase sensitivity, serum proteins are pre-separated by 1D-gel electrophoresis and the gel band corresponding to recombinant human tTF-NGR is investigated. Calibration is performed using the gelseparated pure protein. The method relies on six unique tryptic peptides and is sensitive at the low fmol level.

Eine pseudo-MRM-Methode mittels hochauflösender Massenspektrometrie für den spezifischen Nachweis von truncated tissue factor (tTF; spezifiziert für Tumor-Vaskulatur durch das Peptid GNGRAHA, tTF-NGR) in Serum wird präsentiert. Zur Erhöhung der Nachweisgrenze werden die Serumproteine zunächst durch 1D-Gelelektrophorese getrennt, und nur die Bande, die rekombinantem humanem tTF-NGR entspricht, wird untersucht. Die Kalibrierung wird mit dem gel-getrennten reinen Protein durchgeführt. Die Methode nutzt sechs eindeutige tryptische Peptide und ist bis zu wenigen fmol empfindlich.