Patel, Mikesh;
(2023)
Exploring the untapped reactivity of nitrile reagents for site-selective protein modification.
Doctoral thesis (Ph.D), UCL (University College London).
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Abstract
Novel and effective modes of protein modification are constantly in demand for the development of bioconjugates. Antibody-drug conjugates (ADCs) are a rapidly evolving family of bioconjugates and are based on the well documented monoclonal antibody. These systems provide an excellent model to develop new conjugation techniques due to their diverse and complex nature. The focus of this work investigates the properties and use of the nitrile bis-thiol (NBT) reaction as a new bioconjugation tool. Chapter 2 describes the use of mono-nitriles as a new generation of cysteine-to-lysine (CLT) reagents. New reagent designs have been synthesised and tested on the Fab fragment of the monoclonal antibody; trastuzumab. The CLT product generated amidine conjugates which are stable under a variety of conditions. The designs vary in reactivity via the modulation of the nitrile electron deficiency, eventually leading to a final functional CLT reagent and therefore Fab fragment. Chapter 3 describes the novel chemistry and use of a new, dynamic, bridging reagent in the form of bis-nitriles. These reagents are explored for their disulfide bridging properties and relevant chemistry. The reagents react through a novel chemical cascade resulting in a trapped cyclic product. Functional analogues are then synthesised and tested on the Fab fragment, and later full antibody systems. Alternate nitrile-based bridging reagents are also synthesised and tested to further develop the understanding behind the chemistry and use of such reagents. These bridging reagents were also applied on the protein disulfide isomerase; thioredoxin to realise the potential of these reagents as selective enzymatic inhibitors. The final chapter builds upon the foundation of the NBT reaction with ethanedithiol as a new selective reagent with potential to be developed into a biorthogonal reaction. This can have wide-spread use in areas such as rapid tag systems for use in protein labelling, pull-down assays and potentially more.
| Type: | Thesis (Doctoral) |
|---|---|
| Qualification: | Ph.D |
| Title: | Exploring the untapped reactivity of nitrile reagents for site-selective protein modification |
| Open access status: | An open access version is available from UCL Discovery |
| Language: | English |
| Additional information: | Copyright © The Author 2023. 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 UCL > Provost and Vice Provost Offices > UCL BEAMS UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Maths and Physical Sciences UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Maths and Physical Sciences > Dept of Chemistry |
| URI: | https://discovery-pp.ucl.ac.uk/id/eprint/10184428 |
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