| People | Locations | Statistics |
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| Naji, M. |
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| Motta, Antonella |
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| Aletan, Dirar |
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| Mohamed, Tarek |
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| Ertürk, Emre |
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| Taccardi, Nicola |
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| Kononenko, Denys |
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| Petrov, R. H. | Madrid |
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| Alshaaer, Mazen | Brussels |
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| Bih, L. |
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| Casati, R. |
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| Muller, Hermance |
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| Kočí, Jan | Prague |
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| Šuljagić, Marija |
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| Kalteremidou, Kalliopi-Artemi | Brussels |
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| Azam, Siraj |
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| Ospanova, Alyiya |
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| Blanpain, Bart |
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| Ali, M. A. |
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| Popa, V. |
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| Rančić, M. |
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| Ollier, Nadège |
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| Azevedo, Nuno Monteiro |
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| Landes, Michael |
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| Rignanese, Gian-Marco |
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Desmet, Gert
Vrije Universiteit Brussel
in Cooperation with on an Cooperation-Score of 37%
Topics
Publications (12/12 displayed)
- 2024Detailed analysis of the effective and intra-particle diffusion coefficient of proteins at elevated pressure in columns packed with wide-pore core-shell particlescitations
- 2023Development of a 3D-Printable, Porous, and Chemically Active Material Filled with Silica Particles and its Application to the Fabrication of a Microextraction Devicecitations
- 2022Measurement of the molecular diffusion coefficient and the effective longitudinal diffusion under supercritical fluid chromatography conditions in packed bed columnscitations
- 2022Review of recent insights in the measurement and modelling of the B-term dispersion and related mass transfer properties in liquid chromatographycitations
- 2022Vacuum-driven assembly of electrostatically levitated microspheres on perforated surfacescitations
- 2021Measurement and modelling of the intra-particle diffusion and b-term in reversed-phase liquid chromatographycitations
- 2020Spatial Segregation of Microspheres by Rubbing-Induced Triboelectrification on Patterned Surfacescitations
- 2020A Methodology for the Estimation and Modelling of the Obstruction Factor in the Expression for Mesopore Diffusion in Reversed-Phase Liquid Chromatography Particlescitations
- 2019Study of peak capacities generated by a porous layered radially elongated pillar array column coupled to a nano-LC systemcitations
- 2011High-resolution separations of protein isoforms with liquid chromatography time-of-flight mass spectrometry using polymer monolithic capillary columns
- 2011Ultra-high-resolution separations of intact proteins with LC-TOF-MS using polymer monolithic columns
- 2008Errors involved in the Existing B-term Expressions for the Longitudinal Diffusion in Fully Porous Chromatographic Media. Part II: Experimental data in Packed Columns and Surface Diffusion Measurements
Places of action
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document
Ultra-high-resolution separations of intact proteins with LC-TOF-MS using polymer monolithic columns
Abstract
Top-down approaches to proteomic analyses are attractive, because they have the potential to supply information about the modification status of proteins, including proteolytic processing, oxidative damage, glycosylation, or other forms of post-translational processing. However, several limitations have prevented top-down workflows from becoming widely adopted, in particular (1) fragmentation of proteins, (2) data analysis of the complex MS/MS spectra, (3) chromatographic separation of mixtures of intact proteins, and (4) sensitivity of detection for the intact proteins. To reduce spectral complexity, and also to minimize ion suppression of low abundant species, a high-efficiency separation method is required prior to ESI-MS analysis.In the present contribution, we discuss the separation of intact proteins and protein isoforms arising from various amino-acid modifications employing a poly(styrene-co-divinylbenzene) monolithic capillary column in high-performance liquid chromatography coupled on-line to a time-of-flight mass spectrometry via electrospray interfacing. Using a 250 mm x 0.2 mm monolithic capillary column constructed by in-situ polymerization we have achieved peak capacities of> 600 with a 2h gradient. Detection on a maXis UHR-Qq-TOF of proteins from 200 fmol per protein on columnwith TFA added as ion pairing agent yielded a high signal to noise ratio (11 < S/N < 322) and excellent resolution (res > 30000). Using FA as ion-pairing agent, a 3-fold increase in signal intensity was obtained. The advantages of the combination of ultra-high-resolution chromatography with high-sensitivity high-accuracy MS is demonstrated with the separation of protein isoforms that differ only in their oxidation and glycosylation states.