Site-Specific Radiolabeling of a Human PD-L1 Nanobody via Maleimide-Cysteine Chemistry

Dora Mugoli Chigoho, Quentin Lecocq, Robin Maximilian Awad, Karine Breckpot, Nick Devoogdt, Marleen Keyaerts, Vicky Caveliers, Catarina Xavier, Jessica Bridoux

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11 Citaten (Scopus)
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Samenvatting

Immune checkpoint inhibitors targeting the programmed cell death-1 (PD-1) and its ligand PD-L1 have proven to be efficient cancer therapies in a subset of patients. From all the patients with various cancer types, only 20% have a positive response. Being able to distinguish patients that do express PD-1/PD-L1 from patients that do not allows patients to benefit from a more personalized and efficient treatment of tumor lesion(s). Expression of PD-1 and PD-L1 is typically assessed via immunohistochemical detection in a tumor biopsy. However, this method does not take in account the expression heterogeneity within the lesion, nor the possible metastasis. To visualize whole-body PD-L1 expression by PET imaging, we developed a nanobody-based radio-immunotracer targeting PD-L1 site-specifically labeled with gallium-68. The cysteine-tagged nanobody was site-specifically conjugated with a maleimide (mal)-NOTA chelator and radiolabeling was tested at different nanobody concentrations and temperatures. Affinity and specificity of the tracer, referred to as [68Ga]Ga-NOTA-mal-hPD-L1 Nb, were assayed by surface plasmon resonance and on PD-L1POS or PD-L1NEG 624-MEL cells. Xenografted athymic nude mice bearing 624-MEL PD-L1POS or PD-L1NEG tumors were injected with the tracer and ex vivo biodistribution was performed 1 h 20 min post-injection. Ideal 68Ga-labeling conditions were found at 50 °C for 15 min. [68Ga]Ga-NOTA-mal-hPD-L1 Nb was obtained in 80 ± 5% DC-RCY with a RCP > 99%, and was stable in injection buffer and human serum up to 3 h (>99% RCP). The in vitro characterization showed that the NOTA-functionalized Nb retained its affinity and specificity. Ex vivo biodistribution revealed a tracer uptake of 1.86 ± 0.67% IA/g in the positive tumors compared with 0.42 ± 0.04% IA/g in the negative tumors. Low background uptake was measured in the other organs and tissues, except for the kidneys and bladder, due to the expected excretion route of Nbs. The data obtained show that the site-specific 68Ga-labeled NOTA-mal-hPD-L1 Nb is a promising PET radio-immunotracer due to its ease of production, stability and specificity for PD-L1.

Originele taal-2English
Artikelnummer550
Aantal pagina's11
TijdschriftPharmaceuticals
Volume14
Nummer van het tijdschrift6
DOI's
StatusPublished - 8 jun 2021

Bibliografische nota

Funding Information:
D.M.C. is funded by FWO-Vlaanderen (G085620N). J.B. is funded by the EU H2020 Marie Skłodowska-Curie Actions MSCA ITN PET3D N675417 and by the Scientific Fund W. Gepts UZ Brussel. M.K. is a senior clinical investigator of the Research Foundation Flanders (FWO-Vlaanderen). This research was financially supported by the Belgian Foundation against Cancer (Stichting tegen Kancer, FAF-C/2018/1224); Stand-Up to Cancer (Kom op tegen Kanker), the Flemish cancer society; infrastructure research funding FWO-Vlaanderen (I001618N). Q.L. is funded via an “Emmanuel Vanderscheuren” award and an FWO-SB fellowship (1S24218N). R.M.A. is funded via an FWO-SB fellowship. The research is supported by the Strategic Research Programmes (SRP) of the VUB research council.The authors thank Cindy Peleman, Jan De Jonge and Kevin de Jonghe for their technical assistance.

Funding Information:
Funding: D.M.C. is funded by FWO-Vlaanderen (G085620N). J.B. is funded by the EU H2020 Marie Skłodowska-Curie Actions MSCA ITN PET3D N675417 and by the Scientific Fund W. Gepts UZ Brussel. M.K. is a senior clinical investigator of the Research Foundation Flanders (FWO-Vlaanderen). This research was financially supported by the Belgian Foundation against Cancer (Stichting tegen Kancer, FAF-C/2018/1224); Stand-Up to Cancer (Kom op tegen Kanker), the Flemish cancer society; infrastructure research funding FWO-Vlaanderen (I001618N). Q.L. is funded via an “Emmanuel Vanderscheuren” award and an FWO-SB fellowship (1S24218N). R.M.A. is funded via an FWO-SB fellowship. The research is supported by the Strategic Research Programmes (SRP) of the VUB research council.

Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.

Copyright:
Copyright 2021 Elsevier B.V., All rights reserved.

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