Publications
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Genetic and environmental variation in protein composition of Belgian soy determined with a novel size-exclusion chromatography method KU Leuven Research Institute for Agriculture, Fisheries and Food
Comparison of membrane affinity-based method with size-exclusion chromatography for isolation of exosome-like vesicles from human plasma KU Leuven
Plasma extracellular vesicles (EVs), especially exosome-like vesicles (ELVs), are being increasingly explored as a source of potential noninvasive disease biomarkers. The discovery of blood-based biomarkers associated with ELVs requires methods that isolate high yields of these EVs without significant contamination with highly abundant plasma proteins and lipoproteins. The rising interest in blood-based EV-associated biomarkers has led to the ...
Aqueous size-exclusion chromatographic separations of intact proteins under native conditions: Effect of pressure on selectivity and efficiency Vrije Universiteit Brussel
The selectivity and separation efficiency of aqueous size-exclusion chromatographic separations of intact proteins were assessed for different flow rates, using columns packed with 3 and 5 mu m silica particles containing 150 and 290 angstrom stagnant pores. A mixture of intact proteins with molecular weights ranging between 17 000 and 670 000 Da was used to construct the calibration curves. Both the model fit and the predictive properties, ...
Evaluation of particle and bed integrity of aqueous size-exclusion columns packed with sub-2 µm particles operated at high pressure Vrije Universiteit Brussel
The performance of columns packed with 1.7 µm particles for aqueous size-exclusion chromatography was assessed at high-pressure conditions and linked to particle- and column-bed integrity. Decreasing the particle size from 3.5 µm to 1.7 µm increases the resolution due to the improved mass-transfer characteristics, allowing to significantly speed-up analysis without compromising the selectivity. A sub-minute separation of intact proteins was ...
Advances in native high-performance liquid chromatography and intact mass spectrometry for the characterization of biopharmaceutical products Vrije Universiteit Brussel University of Antwerp
The characterization of biotherapeutics represents a major analytical challenge. This review discusses the current state-of-the-art in analytical technologies to profile biopharma products under native conditions, i.e., the protein three dimensional conformation is maintained during liquid chromatographic analysis. Native liquid-chromatographic modes that are discussed include aqueous size-exclusion chromatography, hydrophobic interaction ...
Development of comprehensive liquid chromatography (LC x LC) approaches for the analysis of complex copolymer structures Ghent University
Knowledge of the exact composition and purity of advanced copolymers is of utmost importance given their influence on the final physical properties. The development of analytical techniques to examine these impurities is therefore crucial with respect of large industrial productions. A comprehensive two-dimensional HPLC separation technique with a slow size exclusion (SEC) separation in the first dimension and a fast reversed phase liquid ...
Impact of extraction and elution media on non-size effects in size exclusion chromatography of proteins KU Leuven
Size exclusion chromatography is extensively used to separate proteins and to determine their apparent molecular weights. It separates proteins based on hydrodynamic volume, but interactions between the chromatography resin and proteins lead to non-size effects. This report discusses the impact of co-solvents [salt, urea, sodium dodecyl sulfate (SDS), dithiothreitol] in extraction media when separating wheat gluten proteins, soy glycinin, bovine ...
Ultrafiltration and size exclusion chromatography combined with asymmetrical-flow field-flow fractionation for the isolation and characterisation of extracellular vesicles from urine University of Antwerp KU Leuven
Extracellular vesicles (EVs) have a great potential in clinical applications. However, their isolation from different bodily fluids and their characterisation are currently not optimal or standardised. Here, we report the results of examining the performance of ultrafiltration combined with size exclusion chromatography (UF-SEC) to isolate EVs from urine. The results reveal that UF-SEC is an efficient method and provides high purity. ...