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

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 sta...

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Autores principales: Chrzastek, Alina, Thanasi, Ioanna A., Irving, James A., Chudasama, Vijay, Baker, James R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Royal Society of Chemistry 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9555722/
https://www.ncbi.nlm.nih.gov/pubmed/36320392
http://dx.doi.org/10.1039/d2sc04531a
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author Chrzastek, Alina
Thanasi, Ioanna A.
Irving, James A.
Chudasama, Vijay
Baker, James R.
author_facet Chrzastek, Alina
Thanasi, Ioanna A.
Irving, James A.
Chudasama, Vijay
Baker, James R.
author_sort Chrzastek, Alina
collection PubMed
description 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.
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spelling pubmed-95557222022-10-31 Dual reactivity disulfide bridging reagents; enabling new approaches to antibody fragment bioconjugation Chrzastek, Alina Thanasi, Ioanna A. Irving, James A. Chudasama, Vijay Baker, James R. Chem Sci Chemistry 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. The Royal Society of Chemistry 2022-09-27 /pmc/articles/PMC9555722/ /pubmed/36320392 http://dx.doi.org/10.1039/d2sc04531a Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by/3.0/
spellingShingle Chemistry
Chrzastek, Alina
Thanasi, Ioanna A.
Irving, James A.
Chudasama, Vijay
Baker, James R.
Dual reactivity disulfide bridging reagents; enabling new approaches to antibody fragment bioconjugation
title Dual reactivity disulfide bridging reagents; enabling new approaches to antibody fragment bioconjugation
title_full Dual reactivity disulfide bridging reagents; enabling new approaches to antibody fragment bioconjugation
title_fullStr Dual reactivity disulfide bridging reagents; enabling new approaches to antibody fragment bioconjugation
title_full_unstemmed Dual reactivity disulfide bridging reagents; enabling new approaches to antibody fragment bioconjugation
title_short Dual reactivity disulfide bridging reagents; enabling new approaches to antibody fragment bioconjugation
title_sort dual reactivity disulfide bridging reagents; enabling new approaches to antibody fragment bioconjugation
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9555722/
https://www.ncbi.nlm.nih.gov/pubmed/36320392
http://dx.doi.org/10.1039/d2sc04531a
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