Fanthom, Thomas Benjamin; (2024) Mechanisms of Therapeutic Protein Aggregation During Peristaltic Pumping. Doctoral thesis (Eng.D), UCL (University College London).

Preview

Text Fanthom_10192749_Thesis.pdf Download (5MB) | Preview

Abstract

This research delves into the stability of recombinant botulinum neurotoxin during bioprocessing, focusing on the challenges posed by turbulent hydrodynamic flow, shear forces, and peristaltic pumping mechanisms. Developing ultra scale down devices to understand the factors governing protein aggregation and monomer loss is essential for the efficient production of therapeutic proteins. In the first study, a comprehensive analysis of flow regimes inside a spinning disc device using computational fluid dynamics gave deeper insights into the flow regimes during operation. Monomer loss and particle formation data gathered from size-exclusion chromatography and backgrounded membrane imaging highlights that turbulent hydrodynamic flow is a significant contributor to -ve BoNT/E protein aggregation. In the second study, a comparative analysis was conducted between a spinning disc device and a closed-loop peristaltic pump device, revealing distinct driving forces governing protein-protein and protein-interface interactions. The spinning disc device's unsuitability as a mimic for peristaltic pumping highlighted the need to explore peristaltic pumping mechanisms further. The third study focused on developing novel methodologies to isolate and assess the impact of various peristaltic pump parameters on monomer loss and protein aggregation. Pump speed, occlusion, and tubing type underscored the crucial role of interfaces in protein adsorption, disruption, aggregation, and particle generation, emphasising their importance in process development and pump setup. Strain on peristaltic pumping via tubing expansion and relaxation caused minor monomer losses, with tubing material influencing the extent of these losses. Solid-solid interface contact was identified as a significant contributor to monomer loss as a function of contact area. In conclusion, this research highlights the solid-solid contact mechanism as the primary driver of protein aggregation during peristaltic pumping. The disruption and the reconstitution of these films create a challenging cycle in peristaltic pump applications. Understanding the underlying mechanisms behind monomer loss and protein aggregation is crucial for the bioprocessing industry to optimise production processes and ensure the stability of therapeutic proteins.

Download activity - last month

Export as JSONCSVXML

Download activity - last 12 months

Export as XMLJSONCSV

Downloads by country - last 12 months

Export as XMLCSVJSON

Archive Staff Only

View Item