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: | |
|---|---|
| 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: | 23.03.2022 |
| 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: | Englisch |
| Format: | PDF document |
| URN: | urn:nbn:de:hbz:6-00269468510 |
| Permalink: | https://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.