Using microdialysis to analyse the passage of monovalent nanobodies through the blood-brain barrier.

TitleUsing microdialysis to analyse the passage of monovalent nanobodies through the blood-brain barrier.
Publication TypeJournal Article
Year of Publication2012
AuthorsCaljon, G., V. Caveliers, T. Lahoutte, B. Stijlemans, G. H. Ghassabeh, J. Van Den Abbeele, I. Smolders, P. De Baetselier, Y. Michotte, S. Muyldermans, S. Magez, and R. Clinckers
JournalBr J Pharmacol
Volume165
Issue7
Pagination2341-53
Date Published2012 Apr
Type of Articlenanobody
ISSN1476-5381
KeywordsAnimals, Antibodies, Protozoan, Blood-Brain Barrier, Male, Microdialysis, Nanostructures, Rats, Rats, Wistar, Technetium Tc 99m Sestamibi, Tomography, Emission-Computed, Single-Photon, Trypanosoma brucei brucei, Trypanosomiasis, African, X-Ray Microtomography
Abstract

BACKGROUND AND PURPOSE: Nanobodies are promising antigen-binding moieties for molecular imaging and therapeutic purposes because of their favourable pharmacological and pharmacokinetic properties. However, the capability of monovalent nanobodies to reach targets in the CNS remains to be demonstrated.EXPERIMENTAL APPROACH: We have assessed the blood-brain barrier permeability of Nb_An33, a nanobody against the Trypanosoma brucei brucei variant-specific surface glycoprotein (VSG). This analysis was performed in healthy rats and in rats that were in the encephalitic stage of African trypanosomiasis using intracerebral microdialysis, single photon emission computed tomography (SPECT) or a combination of both methodologies. This enabled the quantification of unlabelled and (99m) Tc-labelled nanobodies using, respectively, a sensitive VSG-based nanobody-detection elisa, radioactivity measurement in collected microdialysates and SPECT image analysis.KEY RESULTS: The combined read-out methodologies showed that Nb_An33 was detected in the brain of healthy rats following i.v. injection, inflammation-induced damage to the blood-brain barrier, as in the late encephalitic stage of trypanosomiasis, significantly increased the efficiency of passage of the nanobody through this barrier. Complementing SPECT analyses with intracerebral microdialysis improved analysis of brain disposition. There is clear value in assessing penetration of the blood-brain barrier by monovalent nanobodies in models of CNS inflammation. Our data also suggest that rapid clearance from blood might hamper efficient targeting of specific nanobodies to the CNS.CONCLUSIONS AND IMPLICATIONS: Nanobodies can enter the brain parenchyma from the systemic circulation, especially in pathological conditions where the blood-brain barrier integrity is compromised.

DOI10.1111/j.1476-5381.2011.01723.x
Alternate JournalBr. J. Pharmacol.
PubMed ID22013955
PubMed Central IDPMC3413867