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Title: The development of next-generation small volume biophysical screening for the early assessment of monoclonal antibody manufacturability
Author: Joshi, Jay Ketan Kong
ISNI:       0000 0004 9359 2913
Awarding Body: UCL (University College London)
Current Institution: University College London (University of London)
Date of Award: 2020
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The ever-growing need to characterise manufacturability is a key driver for a therapeutic’s success. Early information can reduce costs massively and speed up a product’s delivery to market. Current monoclonal antibody (mAb) methods are limited by the amount of material required and the time it takes to gain valuable information. The formation of aggregates poses a significant hindrance to biopharmaceutical companies. This project aims to develop a novel assay sequence to forecast the manufacturability of a mAb by predicting its propensity to aggregate from low populations. The assay will form part of a biophysical screening process in order to de-risk lead candidates. A panel of six mAb candidates were characterised using currently employed techniques: SEC, DLS and AUC. The limitations were evaluated for each technique. An extensive literature review was conducted to select novel methods to explore as potential replacements for the current methods. After ranking each potential method based on several criteria, fluorescence-based methods were chosen to investigate further. Several fluorescence techniques were evaluated: fluorescence intensity (FLI), red-edge excitation shift (REES) and time-correlated single photon counting (TCSPC). After evaluating each technique, a 14-day time-course study, where samples were isothermally held at elevated temperatures, was carried out to mimic the current lead panel screening. Spearman’s ranking was used to compare the novel techniques to SEC. The earliest time-point at which each technique could detect aggregation of mAbs, formed the basis for a 4-hour time-course study. REES and TCSPC were successful in detecting the aggregation propensity of mAbs. Each technique’s ability to detect concentration-dependent aggregation was also evaluated. TCSPC was successful in identifying the aggregation-prone mAbs from their time-decays, and the impact of different buffers. Finally, a novel assay, using a combination of TCSPC (emission at 330 nm and 395 nm) and REES, was proposed as a method to obtain fast information (<1 minute) on mAbs from low volumes (< 20 μL).
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available