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Bioorthogonal site-selective conjugation of fluorescent dyes to antibodies: method and potential applications

Antibodies are immensely useful tools for biochemical research and have found application in numerous protein detection and purification methods. Moreover, monoclonal antibodies are increasingly utilised as therapeutics or, conjugated to active pharmaceutical ingredients, in targeted chemotherapy. S...

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Autores principales: Grossenbacher, Philipp, Essers, Maria C., Moser, Joël, Singer, Simon A., Häusler, Stephanie, Stieger, Bruno, Rougier, Jean-Sébastien, Lochner, Martin
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/PMC9533196/
https://www.ncbi.nlm.nih.gov/pubmed/36320493
http://dx.doi.org/10.1039/d2ra05580e
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author Grossenbacher, Philipp
Essers, Maria C.
Moser, Joël
Singer, Simon A.
Häusler, Stephanie
Stieger, Bruno
Rougier, Jean-Sébastien
Lochner, Martin
author_facet Grossenbacher, Philipp
Essers, Maria C.
Moser, Joël
Singer, Simon A.
Häusler, Stephanie
Stieger, Bruno
Rougier, Jean-Sébastien
Lochner, Martin
author_sort Grossenbacher, Philipp
collection PubMed
description Antibodies are immensely useful tools for biochemical research and have found application in numerous protein detection and purification methods. Moreover, monoclonal antibodies are increasingly utilised as therapeutics or, conjugated to active pharmaceutical ingredients, in targeted chemotherapy. Several reagents and protocols are reported to synthesise fluorescent antibodies for protein target detection and immunofluorescence applications. However, most of these protocols lead to non-selective conjugation, over-labelling or in the worst case antigen binding site modification. Here, we have used the antibody disulphide cleavage and re-bridging strategy to introduce bright fluorescent dyes without loss of the antibody function. The resulting fluorescent IgG1 type antibodies were shown to be effective imaging tools in western blot and direct immunofluorescence experiments.
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spelling pubmed-95331962022-10-31 Bioorthogonal site-selective conjugation of fluorescent dyes to antibodies: method and potential applications Grossenbacher, Philipp Essers, Maria C. Moser, Joël Singer, Simon A. Häusler, Stephanie Stieger, Bruno Rougier, Jean-Sébastien Lochner, Martin RSC Adv Chemistry Antibodies are immensely useful tools for biochemical research and have found application in numerous protein detection and purification methods. Moreover, monoclonal antibodies are increasingly utilised as therapeutics or, conjugated to active pharmaceutical ingredients, in targeted chemotherapy. Several reagents and protocols are reported to synthesise fluorescent antibodies for protein target detection and immunofluorescence applications. However, most of these protocols lead to non-selective conjugation, over-labelling or in the worst case antigen binding site modification. Here, we have used the antibody disulphide cleavage and re-bridging strategy to introduce bright fluorescent dyes without loss of the antibody function. The resulting fluorescent IgG1 type antibodies were shown to be effective imaging tools in western blot and direct immunofluorescence experiments. The Royal Society of Chemistry 2022-10-05 /pmc/articles/PMC9533196/ /pubmed/36320493 http://dx.doi.org/10.1039/d2ra05580e Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by/3.0/
spellingShingle Chemistry
Grossenbacher, Philipp
Essers, Maria C.
Moser, Joël
Singer, Simon A.
Häusler, Stephanie
Stieger, Bruno
Rougier, Jean-Sébastien
Lochner, Martin
Bioorthogonal site-selective conjugation of fluorescent dyes to antibodies: method and potential applications
title Bioorthogonal site-selective conjugation of fluorescent dyes to antibodies: method and potential applications
title_full Bioorthogonal site-selective conjugation of fluorescent dyes to antibodies: method and potential applications
title_fullStr Bioorthogonal site-selective conjugation of fluorescent dyes to antibodies: method and potential applications
title_full_unstemmed Bioorthogonal site-selective conjugation of fluorescent dyes to antibodies: method and potential applications
title_short Bioorthogonal site-selective conjugation of fluorescent dyes to antibodies: method and potential applications
title_sort bioorthogonal site-selective conjugation of fluorescent dyes to antibodies: method and potential applications
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9533196/
https://www.ncbi.nlm.nih.gov/pubmed/36320493
http://dx.doi.org/10.1039/d2ra05580e
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