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dc.contributor.authorFerreira, Vera FCen
dc.contributor.authorOliveira, Bruno Len
dc.contributor.authorD'Onofrio, Aliceen
dc.contributor.authorFarinha, Carlos Men
dc.contributor.authorGano, Lurdesen
dc.contributor.authorPaulo, Antónioen
dc.contributor.authorLopes Bernardes, Goncaloen
dc.contributor.authorMendes, Filipaen
dc.date.accessioned2020-12-05T00:30:25Z
dc.date.available2020-12-05T00:30:25Z
dc.date.issued2021-01en
dc.identifier.issn1043-1802
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/314771
dc.description.abstractPretargeted imaging has emerged as an effective multi-step strategy aiming to improve imaging contrast and reduce patient radiation exposure through decoupling of the radioactivity from the targeting vector. The inverse electrondemand Diels-Alder (IEDDA) reaction between a trans-cyclooctene (TCO)-conjugated antibody and a labelled tetrazine holds great promise for pretargeted imaging applications due to its bioorthogonality, rapid kinetics under mild conditions, and formation of stable products. Herein, we describe the use of functionalized carbonylacrylic reagents for site-specific incorporation of TCO onto an human epidermal growth factor receptor 2 (HER2) antibody (THIOMAB) containing an engineered unpaired cysteine residue, generating homogeneous conjugates. Precise labelling of THIOMAB-TCO with a fluorescent or radiolabeled tetrazine revealed the potential of the TCO-functionalized antibody for imaging the HER2 after pretargeting in a cellular context in a HER2 positive breast cancer cell line. Control studies with MDA-MD-231 cells, which do not express HER2, further confirmed the target specificity of the modified antibody. THIOMAB-TCO was also evaluated in vivo after pretargeting and subsequent administration of an 111In-labelled tetrazine. Biodistribution studies in breast cancer tumor-bearing mice showed a significant activity accumulation on HER2+ tumors, which was 2.6-fold higher than in HER2– tumors. Additionally, biodistribution studies with THIOMAB without the TCO handle also resulted in a decreased uptake of 111In-DOTA-Tz on HER2+ tumors. Altogether, these results clearly indicate the occurrence of the click reaction at the tumor site, i.e., pretargeting of SK-BR-3 HER2-expressing cells with THIOMAB-TCO and reaction through the TCO moiety present in the antibody. The combined advantages of site-selectivity and stability of TCO tagged-antibodies could allow application of biorthogonal chemistry strategies for pretargeting imaging with minimal side-reactions and background.
dc.format.mediumPrint-Electronicen
dc.languageengen
dc.publisherAmerican Chemical Society
dc.rightsAll rights reserved
dc.rights.uri
dc.titleIn Vivo Pretargeting Based on Cysteine-Selective Antibody Modification with IEDDA Bioorthogonal Handles for Click Chemistry.en
dc.typeArticle
prism.endingPage132
prism.issueIdentifier1en
prism.publicationDate2021en
prism.publicationNameBioconjugate chemistryen
prism.startingPage121
prism.volume32en
dc.identifier.doi10.17863/CAM.61877
dcterms.dateAccepted2020-11-25en
rioxxterms.versionofrecord10.1021/acs.bioconjchem.0c00551en
rioxxterms.versionAM
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserveden
rioxxterms.licenseref.startdate2021-01en
dc.contributor.orcidD'Onofrio, Alice [0000-0001-6740-8607]
dc.contributor.orcidBernardes, Goncalo [0000-0001-6594-8917]
dc.contributor.orcidMendes, Filipa [0000-0003-0646-1687]
dc.identifier.eissn1520-4812
rioxxterms.typeJournal Article/Reviewen
pubs.funder-project-idRoyal Society (URF\R\180019)
pubs.funder-project-idEuropean Commission Horizon 2020 (H2020) Spreading Excellence and Widening Participation (807281)
cam.orpheus.successMon Dec 14 07:30:30 GMT 2020 - Embargo updated*
cam.orpheus.counter1*
rioxxterms.freetoread.startdate2021-12-09


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