Schwach, Jonathan ; Kolobynina, Ksenia ; Brandstetter, Katharina ; Gerlach, Marcus ; Ochtrop, Philipp ; Helma, Jonas ; Hackenberger, Christian P. R. ; Harz, Hartmann ; Cardoso, M. Cristina ; Leonhardt, Heinrich ; Stengl, Andreas (2020)
Site-Specific Antibody Fragment Conjugates for Reversible Staining in Fluorescence Microscopy.
In: Chembiochem : a European journal of chemical biology
doi: 10.1002/cbic.202000727
Artikel, Bibliographie
Dies ist die neueste Version dieses Eintrags.
Kurzbeschreibung (Abstract)
Antibody conjugates have taken a great leap forward as tools in basic and applied molecular life sciences, which 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 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.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2020 |
| Autor(en): | Schwach, Jonathan ; Kolobynina, Ksenia ; Brandstetter, Katharina ; Gerlach, Marcus ; Ochtrop, Philipp ; Helma, Jonas ; Hackenberger, Christian P. R. ; Harz, Hartmann ; Cardoso, M. Cristina ; Leonhardt, Heinrich ; Stengl, Andreas |
| Art des Eintrags: | Bibliographie |
| Titel: | Site-Specific Antibody Fragment Conjugates for Reversible Staining in Fluorescence Microscopy |
| Sprache: | Englisch |
| Publikationsjahr: | 18 November 2020 |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | Chembiochem : a European journal of chemical biology |
| DOI: | 10.1002/cbic.202000727 |
| Zugehörige Links: | |
| Kurzbeschreibung (Abstract): | Antibody conjugates have taken a great leap forward as tools in basic and applied molecular life sciences, which 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 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. |
| ID-Nummer: | pmid:33207032 |
| Fachbereich(e)/-gebiet(e): | 10 Fachbereich Biologie 10 Fachbereich Biologie > Cell Biology and Epigenetics |
| Hinterlegungsdatum: | 24 Nov 2020 07:17 |
| Letzte Änderung: | 08 Jan 2024 07:35 |
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| Suche nach Titel in: | TUfind oder in Google |
Verfügbare Versionen dieses Eintrags
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Site‐Specific Antibody Fragment Conjugates for Reversible Staining in Fluorescence Microscopy. (deposited 05 Jan 2024 13:56)
- Site-Specific Antibody Fragment Conjugates for Reversible Staining in Fluorescence Microscopy. (deposited 24 Nov 2020 07:17) [Gegenwärtig angezeigt]
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