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Title: Application of emerging analytical technologies for characterisation of influenza vaccines
Author: Sofikiti, Antonia
ISNI:       0000 0004 6499 8009
Awarding Body: Newcastle University
Current Institution: University of Newcastle upon Tyne
Date of Award: 2017
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Novartis Vaccines (NV) in Liverpool operates an egg-based, semi-automated, influenza vaccine manufacturing facility with the capability to process a batch per day. This thesis focuses on the Agrippal base product, an inactivated subunit trivalent vaccine. The objectives of this multidisciplinary project included optimisation of assays and introduction of analytical technologies that could potentially facilitate characterisation of viral concentration, particle size and aggregation interactions with a view to promote understanding of product stability and identify opportunities for process optimisation and yield improvements in the Agrippal platform. The first phase of this project focused on methods of nanoparticle analysis that could enable efficient monitoring of viral content from the early stages of product development and throughout the downstream operations. A comparative evaluation between commonly used particle analysers showed that differential centrifugal sedimentation (DCS) is the most suitable method for the analysis of in-process influenza vaccine samples. Novel applications of DCS throughout the manufacturing of egg-based inactivated influenza vaccines were examined and are reported herein. DCS was proven to be a valuable analytical tool by demonstrating high potential for process investigation and monitoring. Key applications include direct determination of viral content in allantoic fluid, viral quantification at whole virus process stages, characterisation of unit operations on a particle based approach and detection and monitoring of the kinetics of aggregation. Furthermore, DCS was specifically employed in a novel study entailing direct determination of influenza virus growth in allantoic fluid upon addition of glucocorticoids and other compounds (hydrocortisone, dexamethasone, corticosterone, AFZ077 and BYF589) into the viral seed inoculum. Additionally, the intricate, multi-step, multi-variable and poorly characterised solubilisation process of the membrane glycoproteins Haemagglutinin (HA) and Neuraminidase (NA) from the surface of influenza virus by the detergent CTAB, as applied in the Agrippal manufacture platform, was comprehensively examined with the aim of enhancing process understanding by identifying the key parameters potentially affecting solubilisation efficiency and product quality. Finally, the development and optimisation of a novel GNA-binding enzyme-linked immunosorbent assay (ELISA) as an alternative replacement to the Single Radial Immunodiffusion (SRID) potency assay for quantification of HA in vaccine samples is described.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (D.Eng.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available