In Vivo Pretargeting Based on Cysteine-Selective Antibody Modification with IEDDA Bioorthogonal Handles for Click Chemistry.
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Authors
Ferreira, Vera FC
Oliveira, Bruno L
Farinha, Carlos M
Gano, Lurdes
Paulo, António
Lopes Bernardes, Goncalo
Publication Date
2021-01Journal Title
Bioconjugate chemistry
ISSN
1043-1802
Publisher
American Chemical Society
Volume
32
Issue
1
Pages
121-132
Language
eng
Type
Article
This Version
AM
Physical Medium
Print-Electronic
Metadata
Show full item recordCitation
Ferreira, V. F., Oliveira, B. L., D'Onofrio, A., Farinha, C. M., Gano, L., Paulo, A., Lopes Bernardes, G., & et al. (2021). In Vivo Pretargeting Based on Cysteine-Selective Antibody Modification with IEDDA Bioorthogonal Handles for Click Chemistry.. Bioconjugate chemistry, 32 (1), 121-132. https://doi.org/10.1021/acs.bioconjchem.0c00551
Abstract
Pretargeted 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.
Sponsorship
Royal Society (URF\R\180019)
European Commission Horizon 2020 (H2020) Spreading Excellence and Widening Participation (807281)
Embargo Lift Date
2021-12-09
Identifiers
External DOI: https://doi.org/10.1021/acs.bioconjchem.0c00551
This record's URL: https://www.repository.cam.ac.uk/handle/1810/314771
Rights
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