Burden, CS;
Jin, J;
Podgornik, A;
Bracewell, DG;
(2012)
A monolith purification process for virus-like particles from yeast homogenate.
Journal of Chromatography B
, 880
(1)
82 - 89.
10.1016/j.jchromb.2011.10.044.
![[thumbnail of 1333870.pdf]](https://discovery.ucl.ac.uk/style/images/fileicons/application_pdf.png) Preview |
PDF
1333870.pdf Download (1MB) |
Abstract
Monoliths are an alternative stationary phase format to conventional particle based media for large biomolecules. Conventional resins suffer from limited capacities and flow rates when used for viruses, virus-like particles (VLP) and other nanoplex materials. The monolith structure provides a more open pore structure to improve accessibility for these materials and better mass transport from convective flow and reduced pressure drops. To examine the performance of this format for bioprocessing we selected the challenging capture of a VLP from clarified yeast homogenate. Using a recombinant Saccharomyces cerevisiae host it was found hydrophobic interaction based separation using a hydroxyl derivatised monolith had the best performance. The monolith was then compared to a known beaded resin method, where the dynamic binding capacity was shown to be three-fold superior for the monolith with equivalent 90% recovery of the VLP. To understand the impact of the crude feed material confocal microscopy was used to visualise lipid contaminants, deriving from the homogenised yeast. It was seen that the lipid formed a layer on top of the column, even after regeneration of the column with isopropanol, resulting in increasing pressure drops with the number of operational cycles. Removal of the lipid pre-column significantly reduces the amount and rate of this fouling process. Using Amberlite/XAD-4 beads around 70% of the lipid was removed, with a loss of VLP around 20%. Applying a reduced lipid feed versus an untreated feed further increased the dynamic binding capacity of the monolith from 0.11 mg/mL column to 0.25 mg/mL column.
| Type: | Article |
|---|---|
| Title: | A monolith purification process for virus-like particles from yeast homogenate. |
| Location: | Netherlands |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1016/j.jchromb.2011.10.044 |
| Publisher version: | http://dx.doi.org/10.1016/j.jchromb.2011.10.044 |
| Language: | English |
| Additional information: | This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
| Keywords: | Ammonium Sulfate, Chromatography, Liquid, Fermentation, Hepatitis B Surface Antigens, Hydrophobic and Hydrophilic Interactions, Lipids, Microscopy, Confocal, Microscopy, Electron, Transmission, Polystyrenes, Porosity, Saccharomyces cerevisiae, Vaccines, Virus-Like Particle |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > UCL BEAMS UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering |
| URI: | https://discovery.ucl.ac.uk/id/eprint/1333870 |
Archive Staff Only
 |
View Item |
