Issue 39, 2022

Dual reactivity disulfide bridging reagents; enabling new approaches to antibody fragment bioconjugation

Abstract

Disulfide bridging, also known as disulfide stapling, is a powerful strategy for the construction of site-selective protein bioconjugates. Here we describe the first examples of a new class of such reagents, containing a ‘stable-labile’ design. These dual-reactive reagents are designed to form a stable bond to one cysteine and a labile bond to the second; resulting in a robust attachment to the protein with one end of the bridge, whilst the other end serves as a reactive handle for subsequent bioconjugation. By incorporating thioesters into these bridges, we demonstrate that they are primed for native chemical ligation (NCL) with N-terminal cysteines; offering an alternative to the requirement for C-terminal thioesters for use in such ligations. Alternatively, the use of hydrazine as the ligating nucleophile enables a separate cargo to be attached to each cysteine residue, which are exploited to insert variably cleavable linkers. These methodologies are demonstrated on an antibody fragment, and serve to expand the scope of disulfide bridging strategies whilst offering a convenient route to the construction of multifunctional antibody fragment conjugates.

Graphical abstract: Dual reactivity disulfide bridging reagents; enabling new approaches to antibody fragment bioconjugation

Supplementary files

Article information

Article type
Edge Article
Submitted
13 Aug 2022
Accepted
16 Sep 2022
First published
27 Sep 2022
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY license

Chem. Sci., 2022,13, 11533-11539

Dual reactivity disulfide bridging reagents; enabling new approaches to antibody fragment bioconjugation

A. Chrzastek, I. A. Thanasi, J. A. Irving, V. Chudasama and J. R. Baker, Chem. Sci., 2022, 13, 11533 DOI: 10.1039/D2SC04531A

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

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