Development & upscaling of 3D printed chromatography columns for virus purification.
| dc.contributor.author | Feast, Sean Ashley | |
| dc.date.accessioned | 2021-06-11T00:19:12Z | |
| dc.date.available | 2021-06-11T00:19:12Z | |
| dc.date.issued | 2021 | en |
| dc.description.abstract | The work presented in this thesis has demonstrated the purification of a range of viruses from both clarified lysate and cell culture using 3D-printed chromatography columns. Previous research and development of 3D-printed columns suggested they would benefit the recovery of virus and virus-like particles used for biopharmaceutical applications such as vaccines and gene therapies. The columns' ability to passage solids while effectively bind and elute a target biological allowed the combination of multiple steps (clarification, filtration, and capture chromatography) in the downstream processing of viruses. This reduction in processing steps and production time should minimize costs for virus production, especially for expensive gene therapies. A triply periodic minimal surface known as the Schoen gyroid formed the cellulose hydrogel- based columns' channel structure. These monolithic-style columns had 300 µm hydraulic diameter channels allowing the passage of solids. Anion exchange (Q or DEAE) or multimodal (hydroxyapatite) ligands were attached to the structure's walls and pores to bind and elute a range of viruses. Both DEAE (229 µeq/mL & 85 mg/mL) and Q (346 µeq/mL & 138 mg/mL) functionalities have comparable ligand densities and BSA static binding capacities compared to bead-based media. Recovery of oncolytic adenovirus (69 ± 6%) and lentivirus (57%) from DEAE columns is high compared with traditional media. Purification of M13 bacteriophage directly from cell culture under comparable conditions to an expanded bed chromatography process showed a high recovery (87.7% ± 5% for 1.49×1011 pfu/mL). Additionally, it was completed three times faster without the difficulties associated with a fluidized bed. This work also acknowledged and examined how this technology could scale from the lab to pilot-size columns capable of processing larger volumes of feedstock. Development of a ‘segmentation method’ alongside commercially available printer advancement led to a single part size increase of 1500%. Fabrication of pilot-sized column (30 cm long by 5 cm diameter, 500 µm channel diameter) is estimated to take 20 hours. | en |
| dc.identifier.uri | https://hdl.handle.net/10092/102023 | |
| dc.identifier.uri | http://dx.doi.org/10.26021/11078 | |
| dc.language | English | |
| dc.language.iso | en | |
| dc.publisher | University of Canterbury | en |
| dc.rights | All Right Reserved | en |
| dc.rights.uri | https://canterbury.libguides.com/rights/theses | en |
| dc.title | Development & upscaling of 3D printed chromatography columns for virus purification. | en |
| dc.type | Theses / Dissertations | en |
| thesis.degree.discipline | Chemical Engineering | en |
| thesis.degree.grantor | University of Canterbury | en |
| thesis.degree.level | Doctoral | en |
| thesis.degree.name | Doctor of Philosophy | en |
| uc.bibnumber | 3047162 | |
| uc.college | Faculty of Engineering | en |