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Unique Benefits of Tumor-Specific Nanobodies for Fluorescence Guided Surgery
Article

The Design and Preclinical Evaluation of a Single-Label Bimodal Nanobody Tracer for Image-Guided Surgery

1
Laboratory for In Vivo Cellular and Molecular Imaging, ICMI-BEFY, Vrije Universiteit Brussel, Laarbeeklaan 103, 1090 Brussels, Belgium
2
Leiden University Medical Center, Interventional Molecular Imaging Laboratory, Department of Radiology, Leiden University, 2311 Leiden, The Netherlands
*
Author to whom correspondence should be addressed.
Academic Editor: Vladimir N. Uversky
Biomolecules 2021, 11(3), 360; https://doi.org/10.3390/biom11030360
Received: 26 January 2021 / Revised: 16 February 2021 / Accepted: 23 February 2021 / Published: 26 February 2021
(This article belongs to the Special Issue The Therapeutic and Diagnostic Potential of Nanobodies)
Intraoperative guidance using targeted fluorescent tracers can potentially provide surgeons with real-time feedback on the presence of tumor tissue in resection margins. To overcome the limited depth penetration of fluorescent light, combining fluorescence with SPECT/CT imaging and/or gamma-ray tracing has been proposed. Here, we describe the design and preclinical validation of a novel bimodal nanobody-tracer, labeled using a “multifunctional single attachment point” (MSAP) label, integrating a Cy5 fluorophore and a diethylenetriaminepentaacetic acid (DTPA) chelator into a single structure. After conjugation of the bimodal MSAP to primary amines of the anti-HER2 nanobody 2Rs15d and 111In-labeling of DTPA, the tracer’s characteristics were evaluated in vitro. Subsequently, its biodistribution and tumor targeting were assessed by SPECT/CT and fluorescence imaging over 24 h. Finally, the tracer’s ability to identify small, disseminated tumor lesions was investigated in mice bearing HER2-overexpressing SKOV3.IP1 peritoneal lesions. [111In]In-MSAP.2Rs15d retained its affinity following conjugation and remained stable for 24 h. In vivo SPECT/CT and fluorescence images showed specific uptake in HER2-overexpressing tumors with low background. High tumor-to-muscle ratios were obtained at 1h p.i. and remained 19-fold on SPECT/CT and 3-fold on fluorescence images over 24 h. In the intraperitoneally disseminated model, the tracer allowed detection of larger lesions via nuclear imaging, while fluorescence enabled accurate removal of submillimeter lesions. Bimodal nuclear/fluorescent nanobody-tracers can thus be conveniently designed by conjugation of a single-molecule MSAP-reagent carrying a fluorophore and chelator for radioactive labeling. Such tracers hold promise for clinical applications. View Full-Text
Keywords: single-domain antibodies; fluorescence-guided surgery; molecular imaging; hybrid imaging single-domain antibodies; fluorescence-guided surgery; molecular imaging; hybrid imaging
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MDPI and ACS Style

Debie, P.; Declerck, N.B.; van Willigen, D.; Huygen, C.M.; De Sloovere, B.; Mateusiak, L.; Bridoux, J.; Puttemans, J.; Devoogdt, N.; van Leeuwen, F.W.B.; Hernot, S. The Design and Preclinical Evaluation of a Single-Label Bimodal Nanobody Tracer for Image-Guided Surgery. Biomolecules 2021, 11, 360. https://doi.org/10.3390/biom11030360

AMA Style

Debie P, Declerck NB, van Willigen D, Huygen CM, De Sloovere B, Mateusiak L, Bridoux J, Puttemans J, Devoogdt N, van Leeuwen FWB, Hernot S. The Design and Preclinical Evaluation of a Single-Label Bimodal Nanobody Tracer for Image-Guided Surgery. Biomolecules. 2021; 11(3):360. https://doi.org/10.3390/biom11030360

Chicago/Turabian Style

Debie, Pieterjan, Noemi B. Declerck, Danny van Willigen, Celine M. Huygen, Bieke De Sloovere, Lukasz Mateusiak, Jessica Bridoux, Janik Puttemans, Nick Devoogdt, Fijs W.B. van Leeuwen, and Sophie Hernot. 2021. "The Design and Preclinical Evaluation of a Single-Label Bimodal Nanobody Tracer for Image-Guided Surgery" Biomolecules 11, no. 3: 360. https://doi.org/10.3390/biom11030360

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