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2020-11-18Zeitschriftenartikel DOI: 10.1002/cbic.202000727
Site‐Specific Antibody Fragment Conjugates for Reversible Staining in Fluorescence Microscopy
dc.contributor.authorSchwach, Jonathan
dc.contributor.authorKolobynina, Ksenia
dc.contributor.authorBrandstetter, Katharina
dc.contributor.authorGerlach, Marcus
dc.contributor.authorOchtrop, Philipp
dc.contributor.authorHelma, Jonas
dc.contributor.authorHackenberger, Christian
dc.contributor.authorHarz, Hartmann
dc.contributor.authorCardoso, M. Cristina
dc.contributor.authorLeonhardt, Heinrich
dc.contributor.authorStengl, Andreas
dc.date.accessioned2021-04-30T08:45:32Z
dc.date.available2021-04-30T08:45:32Z
dc.date.issued2020-11-18none
dc.date.updated2021-02-15T09:40:01Z
dc.identifier.urihttp://edoc.hu-berlin.de/18452/23525
dc.description.abstractAntibody 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.eng
dc.description.sponsorshipTub-tag® technology
dc.description.sponsorshipDeutsche Forschungsgemeinschaft http://dx.doi.org/10.13039/501100001659
dc.description.sponsorshipGerman Research Foundation
dc.description.sponsorshipLeibniz Association http://dx.doi.org/10.13039/501100001664
dc.description.sponsorshipLeibniz Wettbewerb
dc.language.isoengnone
dc.publisherHumboldt-Universität zu Berlin
dc.rights(CC BY 4.0) Attribution 4.0 Internationalger
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subjectantibody conjugateseng
dc.subjectimmunofluorescence stainingeng
dc.subjectnanobodieseng
dc.subjectpeptide nucleic acidseng
dc.subjectsuper-resolution microscopyeng
dc.subjectTub-tag labellingeng
dc.subject.ddc540 Chemie und zugeordnete Wissenschaftennone
dc.subject.ddc572 Biochemienone
dc.titleSite‐Specific Antibody Fragment Conjugates for Reversible Staining in Fluorescence Microscopynone
dc.typearticle
dc.identifier.urnurn:nbn:de:kobv:11-110-18452/23525-5
dc.identifier.doi10.1002/cbic.202000727none
dc.identifier.doihttp://dx.doi.org/10.18452/22853
dc.type.versionpublishedVersionnone
local.edoc.container-titleChemBioChem : a European journal of chemical biologynone
local.edoc.pages6none
local.edoc.type-nameZeitschriftenartikel
local.edoc.institutionMathematisch-Naturwissenschaftliche Fakultätnone
local.edoc.container-typeperiodical
local.edoc.container-type-nameZeitschrift
local.edoc.container-publisher-nameWiley-VCHnone
local.edoc.container-publisher-placeWeinheimnone
local.edoc.container-volume22none
local.edoc.container-issue7none
local.edoc.container-firstpage1205none
local.edoc.container-lastpage1209none
dc.description.versionPeer Reviewednone
local.edoc.affiliationSchwach, Jonathan: Ludwig-Maximilians-Universität München Department of Biology II, Human Biology and BioImaging 82152 Planegg-Martinsried Germanynone
local.edoc.affiliationKolobynina, Ksenia: Technical University of Darmstadt Department of Biology, Cell Biology and Epigenetics Schnittspahnstr. 10 64287 Darmstadt Germanynone
local.edoc.affiliationBrandstetter, Katharina: Ludwig-Maximilians-Universität München Department of Biology II, Human Biology and BioImaging 82152 Planegg-Martinsried Germanynone
local.edoc.affiliationGerlach, Marcus: Tubulis GmbH, BioSysM Butenandtstrasse 1 81377 Munich Germanynone
local.edoc.affiliationOchtrop, Philipp: Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP) Department Chemical Biology Robert-Rössle-Strasse 10 13125 Berlin Germanynone
local.edoc.affiliationHelma, Jonas: Tubulis GmbH, BioSysM Butenandtstrasse 1 81377 Munich Germanynone
local.edoc.affiliationHackenberger, Christian P. R.: Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP) Department Chemical Biology Robert-Rössle-Strasse 10 13125 Berlin Germany; Humboldt Universität zu Berlin, Department of Chemistry, Brook-Taylor-Strasse 2, 12489 Berlin (Germany)none
local.edoc.affiliationHarz, Hartmann: Ludwig-Maximilians-Universität München Department of Biology II, Human Biology and BioImaging 82152 Planegg-Martinsried Germanynone
local.edoc.affiliationCardoso, M. Cristina: Technical University of Darmstadt Department of Biology, Cell Biology and Epigenetics Schnittspahnstr. 10 64287 Darmstadt Germanynone
local.edoc.affiliationLeonhardt, Heinrich: Ludwig-Maximilians-Universität München Department of Biology II, Human Biology and BioImaging 82152 Planegg-Martinsried Germanynone
local.edoc.affiliationStengl, Andreas: Ludwig-Maximilians-Universität München Department of Biology II, Human Biology and BioImaging 82152 Planegg-Martinsried Germanynone

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