Engineering the sialome of mammalian cells with sialic acid mimetics

Mammalian glycans show a diversity in sialic acid capping, constituting the sialome. Sialic acids can be extensively modified chemically, yielding sialic acid mimetics (SAMs). Here, we present a protocol for detecting and quantifying incorporative SAMs using microscopy and flow cytometry, respective...

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Detalles Bibliográficos
Autores principales: Hornikx, Daniël L.A.H., Visser, Eline A., Psomiadou, Venetia, Büll, Christian, Boltje, Thomas J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10511862/
https://www.ncbi.nlm.nih.gov/pubmed/37389993
http://dx.doi.org/10.1016/j.xpro.2023.102330
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author Hornikx, Daniël L.A.H.
Visser, Eline A.
Psomiadou, Venetia
Büll, Christian
Boltje, Thomas J.
author_facet Hornikx, Daniël L.A.H.
Visser, Eline A.
Psomiadou, Venetia
Büll, Christian
Boltje, Thomas J.
author_sort Hornikx, Daniël L.A.H.
collection PubMed
description Mammalian glycans show a diversity in sialic acid capping, constituting the sialome. Sialic acids can be extensively modified chemically, yielding sialic acid mimetics (SAMs). Here, we present a protocol for detecting and quantifying incorporative SAMs using microscopy and flow cytometry, respectively. We detail steps for linking SAMS to proteins with western blotting. Lastly, we detail procedures for incorporative or inhibitory SAMs and how SAMs can be used for the on-cell synthesis of high-affinity Siglec ligands. For complete details on the use and execution of this protocol, please refer to Büll et al.(1) and Moons et al.(2)
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spelling pubmed-105118622023-09-22 Engineering the sialome of mammalian cells with sialic acid mimetics Hornikx, Daniël L.A.H. Visser, Eline A. Psomiadou, Venetia Büll, Christian Boltje, Thomas J. STAR Protoc Protocol Mammalian glycans show a diversity in sialic acid capping, constituting the sialome. Sialic acids can be extensively modified chemically, yielding sialic acid mimetics (SAMs). Here, we present a protocol for detecting and quantifying incorporative SAMs using microscopy and flow cytometry, respectively. We detail steps for linking SAMS to proteins with western blotting. Lastly, we detail procedures for incorporative or inhibitory SAMs and how SAMs can be used for the on-cell synthesis of high-affinity Siglec ligands. For complete details on the use and execution of this protocol, please refer to Büll et al.(1) and Moons et al.(2) Elsevier 2023-06-28 /pmc/articles/PMC10511862/ /pubmed/37389993 http://dx.doi.org/10.1016/j.xpro.2023.102330 Text en © 2023 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Hornikx, Daniël L.A.H.
Visser, Eline A.
Psomiadou, Venetia
Büll, Christian
Boltje, Thomas J.
Engineering the sialome of mammalian cells with sialic acid mimetics
title Engineering the sialome of mammalian cells with sialic acid mimetics
title_full Engineering the sialome of mammalian cells with sialic acid mimetics
title_fullStr Engineering the sialome of mammalian cells with sialic acid mimetics
title_full_unstemmed Engineering the sialome of mammalian cells with sialic acid mimetics
title_short Engineering the sialome of mammalian cells with sialic acid mimetics
title_sort engineering the sialome of mammalian cells with sialic acid mimetics
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10511862/
https://www.ncbi.nlm.nih.gov/pubmed/37389993
http://dx.doi.org/10.1016/j.xpro.2023.102330
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