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 (2024)
Site‐Specific Antibody Fragment Conjugates for Reversible Staining in Fluorescence Microscopy.
In: ChemBioChem, 2020, 22 (7)
doi: 10.26083/tuprints-00017776
Artikel, Zweitveröffentlichung, Verlagsversion
 | Es ist eine neuere Version dieses Eintrags verfügbar. |
Kurzbeschreibung (Abstract)
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.
| Typ des Eintrags: | Artikel | ||||
|---|---|---|---|---|---|
| Erschienen: | 2024 | ||||
| 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: | Zweitveröffentlichung | ||||
| Titel: | Site‐Specific Antibody Fragment Conjugates for Reversible Staining in Fluorescence Microscopy | ||||
| Sprache: | Englisch | ||||
| Publikationsjahr: | 5 Januar 2024 | ||||
| Ort: | Darmstadt | ||||
| Publikationsdatum der Erstveröffentlichung: | 2020 | ||||
| Ort der Erstveröffentlichung: | Weinheim | ||||
| Verlag: | Wiley-VCH | ||||
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | ChemBioChem | ||||
| Jahrgang/Volume einer Zeitschrift: | 22 | ||||
| (Heft-)Nummer: | 7 | ||||
| DOI: | 10.26083/tuprints-00017776 | ||||
| URL / URN: | https://tuprints.ulb.tu-darmstadt.de/17776 | ||||
| Zugehörige Links: | |||||
| Herkunft: | Zweitveröffentlichung DeepGreen | ||||
| Kurzbeschreibung (Abstract): | 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. |
||||
| Alternatives oder übersetztes Abstract: |
|
||||
| Freie Schlagworte: | antibody conjugates, immunofluorescence staining, nanobodies, peptide nucleic acids, super-resolution microscopy, Tub-tag labelling | ||||
| Status: | Verlagsversion | ||||
| URN: | urn:nbn:de:tuda-tuprints-177764 | ||||
| Zusätzliche Informationen: | This article also appears in: Frontiers in Medicinal Chemistry 2022 SPP 1623 - Chemoselective reactions for the synthesis and application of functional proteins |
||||
| Sachgruppe der Dewey Dezimalklassifikatin (DDC): | 500 Naturwissenschaften und Mathematik > 540 Chemie 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie |
||||
| Fachbereich(e)/-gebiet(e): | 10 Fachbereich Biologie 10 Fachbereich Biologie > Cell Biology and Epigenetics |
||||
| Hinterlegungsdatum: | 05 Jan 2024 13:56 | ||||
| Letzte Änderung: | 08 Jan 2024 07:33 | ||||
| PPN: | |||||
| Zugehörige Links: | |||||
| Export: | |||||
| Suche nach Titel in: | TUfind oder in Google | ||||
Verfügbare Versionen dieses Eintrags
- Site‐Specific Antibody Fragment Conjugates for Reversible Staining in Fluorescence Microscopy. (deposited 05 Jan 2024 13:56) [Gegenwärtig angezeigt]
 |
Frage zum Eintrag |
Optionen (nur für Redakteure)
 |
Redaktionelle Details anzeigen |
Drucken
Impressum