Pandya, Akash Rajnikant;
(2022)
Mapping the interaction between excipients with proteins in formulations by computational modelling and NMR.
Doctoral thesis (Ph.D), UCL (University College London).
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Abstract
Monoclonal antibody (mAb) based products are a dominant class of biopharmaceuticals. There is a need for the preferred liquid aqueous form to achieve a long-shelf life of up to 2 years at 4 ℃ in storage. Aggregation has made designing the optimal formulation significantly challenging. This study aims to characterise excipient-protein interactions using a combination of experimental (Tm and STD NMR) and computational (molecular docking and MD simulations) methods. Overall, the presence of excipients used in this study improved the Tm of Fab A33. Interactions between the different excipients and Fab fragment were probed using molecular docking. Multiple interaction hotspots for each excipient were found suggesting the non-specific binding nature of excipients. Results from MD simulations on citrate-glycine-Fab formulations revealed that glycine was preferentially included at lower concentrations (GLY10 and GLY20) and excluded at higher concentrations (GLY30 to GLY60) from the local domain of Fab A33. The largest increase in Tm and decrease in RMSF of Fab was found at GLY10. The gain in citrate interactions was thought to contribute to these trends. Increased preferential exclusion and the displacement of citrate from the Fab surface was found to drive the increase (slower) in the Tm and the increase in RMSF. The increase in exclusion of glycine, no further citrate displacement and monomeric glycine binding was thought to be responsible for the decrease in RMSF at GLY50 and GLY60. Overall, the Tm was found to be affected by specific interactions between monomer glycine and Fab. The ΔRMSF also correlated well with monomeric glycine, leading to the conclusion, that glycine makes Fab A33 more dynamic, and however this does not translate into stability. Overall, these results provide valuable insights into excipient-protein interactions, which can be used to inform the design of formulations.
| Type: | Thesis (Doctoral) |
|---|---|
| Qualification: | Ph.D |
| Title: | Mapping the interaction between excipients with proteins in formulations by computational modelling and NMR |
| Open access status: | An open access version is available from UCL Discovery |
| Language: | English |
| Additional information: | Copyright © The Author 2022. Original content in this thesis is licensed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0) Licence (https://creativecommons.org/licenses/by-nc/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 > 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 UCL > Provost and Vice Provost Offices > UCL BEAMS UCL |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10143029 |
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