Single-Payload Bioconjugation to Native Antibodies And A General Method for the Self-Immolative Release of Amide-Containing Molecules
type: CreativeWork
description: Antibody-drug conjugates (ADCs) have emerged as a powerful form of targeted cancer therapy that can deliver drugs with a high level of selectivity towards a specific cell type, reducing off-target effects and increasing the therapeutic window compared to small molecule therapeutics. However, creating ADCs that are stable, homogeneous, and with controlled drug-to-antibody ratio (DAR) remains a significant challenge. Chapter 1 describes the bioconjugation optimisation of the third generation of tetra-divinylpyrimidine (TetraDVP) linkers. These linkers allow for the creation of single-payload antibody conjugates by re-bridging all four interchain disulphide bonds in IgG1 antibodies with a single construct. Unlike earlier generations, the TetraDVPs used in this work contain the payload before the bioconjugation occurs; this means the entire construct is added in a single step ‘all-at-once’, reducing the antibody manipulation required. The optimised bioconjugation includes a modular reagent addition cycle approach that enabled constructs with varied payloads to be conjugated by >93% in all cases by simply adapting the number of addition cycles. This enabled the creation of protein catcher SpyTag, cytotoxic MMAE, and fluorophore BODIPY single-payload conjugates of trastuzumab. Flow cytometry and SpyTag-SpyCatcher studies carried out by Claudia Driscoll, Howarth group, showed that the ‘all-at-once’ TetraDVP concept can be used to make single-payload antibody constructs that maintain the activity and binding of the antibody and the payload. The controlled release of drug molecules is of interest across various chemical fields. It allows for the masking of a molecule's properties and precise deployment upon a single controllable release event. Although numerous methodologies have been developed for amines, alcohols, and thiols, approaches for utilising amides as payload-release handles have seldom been reported despite the prevalence of amides in therapeutic compounds and ma…
abstract: Antibody-drug conjugates (ADCs) have emerged as a powerful form of targeted cancer therapy that can deliver drugs with a high level of selectivity towards a specific cell type, reducing off-target effects and increasing the therapeutic window compared to small molecule therapeutics. However, creating ADCs that are stable, homogeneous, and with controlled drug-to-antibody ratio (DAR) remains a significant challenge. Chapter 1 describes the bioconjugation optimisation of the third generation of tetra-divinylpyrimidine (TetraDVP) linkers. These linkers allow for the creation of single-payload antibody conjugates by re-bridging all four interchain disulphide bonds in IgG1 antibodies with a single construct. Unlike earlier generations, the TetraDVPs used in this work contain the payload before the bioconjugation occurs; this means the entire construct is added in a single step ‘all-at-once’, reducing the antibody manipulation required. The optimised bioconjugation includes a modular reagent addition cycle approach that enabled constructs with varied payloads to be conjugated by >93% in all cases by simply adapting the number of addition cycles. This enabled the creation of protein catcher SpyTag, cytotoxic MMAE, and fluorophore BODIPY single-payload conjugates of trastuzumab. Flow cytometry and SpyTag-SpyCatcher studies carried out by Claudia Driscoll, Howarth group, showed that the ‘all-at-once’ TetraDVP concept can be used to make single-payload antibody constructs that maintain the activity and binding of the antibody and the payload. The controlled release of drug molecules is of interest across various chemical fields. It allows for the masking of a molecule's properties and precise deployment upon a single controllable release event. Although numerous methodologies have been developed for amines, alcohols, and thiols, approaches for utilising amides as payload-release handles have seldom been reported despite the prevalence of amides in therapeutic compounds and ma…
author: Wharton, Thomas
language: eng
keywords: Antibody-Drug Conjugates, Chemical Biology, Organic Chemistry
license: https://www.repository.cam.ac.uk/bitstreams/0c0f31ee-0c79-490a-b38d-d6a8191a0e47/download
license: https://creativecommons.org/licenses/by/4.0/
date published: 2024-09-25
date modified: 2026-07-24T01:33:15Z
provider: Cambridge University
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: University of Cambridge

PropertyValue
property ID: DOI
value: 10.17863/CAM.114785