Preclinical screening of anti-HER2 nanobodies for molecular imaging of breast cancer.

TitlePreclinical screening of anti-HER2 nanobodies for molecular imaging of breast cancer.
Publication TypeJournal Article
Year of Publication2011
AuthorsVaneycken, I., N. Devoogdt, N. Van Gassen, C. Vincke, C. Xavier, U. Wernery, S. Muyldermans, T. Lahoutte, and V. Caveliers
JournalFASEB J
Volume25
Issue7
Pagination2433-46
Date Published2011 Jul
ISSN1530-6860
KeywordsAnimals, Antibody Specificity, Breast Neoplasms, Camelids, New World, Cell Line, Tumor, CHO Cells, Cricetinae, Cricetulus, Female, Humans, Mammary Neoplasms, Experimental, Mice, Mice, Nude, Molecular Imaging, Peptide Library, Receptor, erbB-2, Single-Chain Antibodies, Technetium, Tissue Distribution, Tomography, Emission-Computed, Single-Photon, Transplantation, Heterologous, X-Ray Microtomography
Abstract

Accurate determination of tumor human epidermal growth factor receptor 2 (HER2)-status in breast cancer patients is possible via noninvasive imaging, provided adequate tracers are used. In this study, we describe the generation of a panel of 38 nanobodies, small HER2-binding fragments that are derived from heavy-chain-only antibodies raised in an immunized dromedary. In search of a lead compound, a subset of nanobodies was biochemically characterized in depth and preclinically tested for use as tracers for imaging of xenografted tumors. The selected compound, 2Rs15d, was found to be stable and to interact specifically with HER2 recombinant protein and HER2-expressing cells in ELISA, surface plasmon resonance, flow cytometry, and radioligand binding studies with low nanomolar affinities, and did not compete with anti-HER2 therapeutic antibodies trastuzumab and pertuzumab. Single-photon-emission computed tomography (SPECT) imaging quantification and biodistribution analyses showed that (99m)Tc-labeled 2Rs15d has a high tumor uptake in 2 HER2(+) tumor models, fast blood clearance, low accumulation in nontarget organs except kidneys, and high concomitant tumor-to-blood and tumor-to-muscle ratios at 1 h after intravenous injection. These values were dramatically lower for an irrelevant control (99m)Tc-nanobody and for (99m)Tc-2Rs15d targeting a HER2(-) tumor.

DOI10.1096/fj.10-180331
Alternate JournalFASEB J.
PubMed ID21478264