Joshi, Jay Ketan Kong;
(2020)
The development of next-generation small volume biophysical screening for the early assessment of monoclonal antibody manufacturability.
Doctoral thesis (Ph.D), UCL (University College London).
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Abstract
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).
| Type: | Thesis (Doctoral) |
|---|---|
| Qualification: | Ph.D |
| Title: | The development of next-generation small volume biophysical screening for the early assessment of monoclonal antibody manufacturability |
| Event: | UCL |
| Open access status: | An open access version is available from UCL Discovery |
| Language: | English |
| Additional information: | Copyright © The Author 2020. Original content in this thesis is licensed under the terms of the Creative Commons Attribution 4.0 International (CC BY 4.0) Licence (https://creativecommons.org/licenses/by/4.0/). Any third-party copyright material present remains the property of its respective owner(s) and is licensed under its existing terms. Access may initially be restricted at the author’s request. |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices 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/10100186 |
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