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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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Royal Society of Chemistry
2022
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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. |
format | Online Article Text |
id | pubmed-9555722 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | The Royal Society of Chemistry |
record_format | MEDLINE/PubMed |
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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