A Novel Universal Detection Agent for Time-Gated Luminescence Bioimaging

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dc.contributor.author Sayyadi, N en
dc.contributor.author Care, A en
dc.contributor.author Connally, RE en
dc.contributor.author Try, AC en
dc.contributor.author Bergquist, Peter en
dc.contributor.author Sunna, A en
dc.date.accessioned 2016-07-11T05:08:56Z en
dc.date.issued 2016-01 en
dc.identifier.citation Scientific Reports, 2016, 6 pp. 1 - 9 en
dc.identifier.issn 2045-2322 en
dc.identifier.uri http://hdl.handle.net/2292/29366 en
dc.description.abstract Luminescent lanthanide chelates have been used to label antibodies in time-gated luminescence (TGL) bioimaging. However, it is a challenging task to label directly an antibody with lanthanide-binding ligands and achieve control of the target ligand/protein ratios whilst ensuring that affinity and avidity of the antibody remain uncompromised. We report the development of a new indirect detection reagent to label antibodies with detectable luminescence that circumvents this problem by labelling available lysine residues in the linker portion of the recombinant fusion protein Linker-Protein G (LPG). Succinimide-activated lanthanide chelating ligands were attached to lysine residues in LPG and Protein G (without Linker) and the resulting Luminescence-Activating (LA-) conjugates were compared for total incorporation and conjugation efficiency. A higher and more efficient incorporation of ligands at three different molar ratios was observed for LPG and this effect was attributed to the presence of eight readily available lysine residues in the linker region of LPG. These Luminescence-Activating (LA-) complexes were subsequently shown to impart luminescence (upon formation of europium(III) complexes) to cell-specific antibodies within seconds and without the need for any complicated bioconjugation procedures. The potential of this technology was demonstrated by direct labelling of Giardia cysts and Cryptosporidium oocysts in TGL bioimaging. en
dc.description.uri http://www.nature.com/srep/ en
dc.format.medium Electronic en
dc.language eng en
dc.publisher Nature Publishing Group en
dc.relation.ispartofseries Scientific Reports en
dc.rights Items in ResearchSpace are protected by copyright, with all rights reserved, unless otherwise indicated. Previously published items are made available in accordance with the copyright policy of the publisher. Details obtained from http://www.sherpa.ac.uk/romeo/issn/2045-2322/ en
dc.rights.uri https://researchspace.auckland.ac.nz/docs/uoa-docs/rights.htm en
dc.rights.uri https://creativecommons.org/licenses/by/4.0/ en
dc.title A Novel Universal Detection Agent for Time-Gated Luminescence Bioimaging en
dc.type Journal Article en
dc.identifier.doi 10.1038/srep27564 en
pubs.begin-page 1 en
pubs.volume 6 en
dc.description.version VoR – Version of Record en
dc.rights.holder Copyright: Nature Publishing Group en
dc.identifier.pmid 27282464 en
pubs.author-url http://www.nature.com/articles/srep27564 en
pubs.end-page 9 en
dc.rights.accessrights http://purl.org/eprint/accessRights/OpenAccess en
pubs.subtype Article en
pubs.elements-id 531496 en
dc.identifier.eissn 2045-2322 en
pubs.number 27564 en
pubs.record-created-at-source-date 2016-07-11 en
pubs.dimensions-id 27282464 en

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