Site-Specific Antibody Fragment Conjugates for Reversible Staining in Fluorescence Microscopy.
Alkynes
/ chemistry
Azides
/ chemistry
Catalysis
Cell Line
Copper
/ chemistry
Cycloaddition Reaction
DNA
/ chemistry
Green Fluorescent Proteins
/ chemistry
Humans
Immunoconjugates
/ chemistry
Immunoglobulin Fragments
/ chemistry
Microscopy, Fluorescence
Peptide Nucleic Acids
/ chemistry
Single-Domain Antibodies
/ chemistry
Tub-tag labelling
antibody conjugates
immunofluorescence staining
nanobodies
peptide nucleic acids
super-resolution microscopy
Journal
Chembiochem : a European journal of chemical biology
ISSN: 1439-7633
Titre abrégé: Chembiochem
Pays: Germany
ID NLM: 100937360
Informations de publication
Date de publication:
06 04 2021
06 04 2021
Historique:
revised:
17
11
2020
received:
21
10
2020
pubmed:
19
11
2020
medline:
18
11
2021
entrez:
18
11
2020
Statut:
ppublish
Résumé
Antibody conjugates have taken a great leap forward as tools in basic and applied molecular life sciences that was enabled by the development of chemoselective reactions for the site-specific modification of proteins. Antibody-oligonucleotide conjugates combine the antibody's target specificity with the reversible, sequence-encoded binding properties of oligonucleotides like DNAs or peptide nucleic acids (PNAs), allowing sequential imaging of large numbers of targets in a single specimen. In this report, we use the Tub-tag® technology in combination with Cu-catalyzed azide-alkyne cycloaddition for the site-specific conjugation of single DNA and PNA strands to an eGFP-binding nanobody. We show binding of the conjugate to recombinant eGFP and subsequent sequence-specific annealing of fluorescently labelled imager strands. Furthermore, we reversibly stain eGFP-tagged proteins in human cells, thus demonstrating the suitability of our conjugation strategy to generate antibody-oligonucleotides for reversible immunofluorescence imaging.
Identifiants
pubmed: 33207032
doi: 10.1002/cbic.202000727
pmc: PMC8048457
doi:
Substances chimiques
Alkynes
0
Azides
0
Immunoconjugates
0
Immunoglobulin Fragments
0
Peptide Nucleic Acids
0
Single-Domain Antibodies
0
enhanced green fluorescent protein
0
Green Fluorescent Proteins
147336-22-9
Copper
789U1901C5
DNA
9007-49-2
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1205-1209Subventions
Organisme : Tub-tag® technology
Organisme : Deutsche Forschungsgemeinschaft
Organisme : German Research Foundation
ID : SPP1623
Organisme : German Research Foundation
ID : 4468/9-1
Organisme : German Research Foundation
ID : 9-2
Organisme : German Research Foundation
ID : 10-2
Organisme : German Research Foundation
ID : 721/13-1
Organisme : German Research Foundation
ID : 13-2
Organisme : German Research Foundation
ID : 198/8-1
Organisme : German Research Foundation
ID : 8-2
Organisme : German Research Foundation
ID : GRK1657/TP1B
Organisme : German Research Foundation
ID : GRK1721
Organisme : Leibniz Association
Organisme : Leibniz Wettbewerb
ID : T18/2017
Informations de copyright
© 2020 The Authors. ChemBioChem published by Wiley-VCH GmbH.
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