Generation and characterization of nanobodies targeting PSMA for molecular imaging of prostate cancer.

TitleGeneration and characterization of nanobodies targeting PSMA for molecular imaging of prostate cancer.
Publication TypeJournal Article
Year of Publication2014
AuthorsEvazalipour, M., M. D'huyvetter, B. Soltani Tehrani, M. Abolhassani, K. Omidfar, S. Abdoli, R. Arezumand, H. Morovvati, T. Lahoutte, S. Muyldermans, and N. Devoogdt
JournalContrast Media Mol Imaging
Volume9
Issue3
Pagination211-20
Date Published2014 May-Jun
ISSN1555-4317
Abstract

Nanobodies show attractive characteristics for tumor targeting in cancer diagnosis and therapy. A radiolabeled nanobody binding the prostate-specific membrane antigen (PSMA) could offer a noninvasive strategy to select prostate cancer patients eligible for PSMA-targeted therapies. We here describe the generation, production and in vivo evaluation of anti-PSMA nanobodies. Nanobodies were derived from heavy-chain-only antibodies, raised in immunized dromedaries. Binding characteristics were evaluated through ELISA and flow cytometry. Selected nanobodies were radiolabeled with (99m) Tc at their hexahistidine tail, after which cell binding capacity and internalization were evaluated on PSMA(pos) LNCaP and PSMA(neg) PC3 cell lines. In vivo tumor targeting was analyzed in both LNCaP and PC3 xenografted mice through SPECT/microCT and tissue sampling. A panel of 72 generated clones scored positive on ELISA, all contributing to three nanobody groups, of which group 3 dominated with 70 clones. ELISA and FACS analysis led to the selection of two dominant nanobodies. (99m) Tc-labeled PSMA6 and PSMA30 both showed specific binding on LNCAP cells, but not on PC3 cells. (99m) Tc-PSMA30 internalized significantly more in LNCaP cells compared to (99m) Tc-PSMA6. Higher absolute tumor uptake and tumor-to-normal organ ratios were observed for (99m) Tc-PSMA30 compared with (99m) Tc-PSMA6 and a (99m) Tc-control nanobody in LNCaP but not in PC3 tumor-bearing mice. PSMA30 nanobody has improved targeting characteristics both in vitro as well as in vivo compared with PSMA6 and the control nanobody, and was therefore selected as our in-house-developed lead compound for PSMA targeting.

DOI10.1002/cmmi.1558
Alternate JournalContrast Media Mol Imaging
PubMed ID24700748
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