Selection of single domain anti-transferrin receptor antibodies for blood-brain barrier transcytosis using a neurotensin based assay and histological assessment of target engagement in a mouse model of Alzheimer’s related amyloid-beta pathology

The blood-brain barrier (BBB) presents a major obstacle in developing specific diagnostic imaging agents for many neurological disorders. In this study we aimed to generate single domain anti-mouse transferrin receptor antibodies (anti-mTfR VHHs) to mediate BBB transcytosis as components of novel MR...

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Bibliographic Details
Published in:PloS one 2022-10, Vol.17 (10), p.e0276107-e0276107
Main Authors: Su, Shiran, Esparza, Thomas J, Brody, David L
Format: Article
Language:English
Subjects:
Affinity
Alzheimer's disease
Antibodies
Binding
Biology and Life Sciences
Blood-brain barrier
Body weight
Care and treatment
Contrast agents
Contrast media
Cytotoxicity
Dimers
Dissociation
Dyes
Fluorescent dyes
Fluorescent indicators
Health aspects
Hypothalamus
Hypothermia
Immunization
Injection
Intravenous administration
Labelling
Magnetic resonance imaging
Medical imaging
Medicine and Health Sciences
Molecular weight
Monoclonal antibodies
Mutants
Nanoparticles
Neuroimaging
Neurological diseases
Neurotensin
Pathology
Patient outcomes
Phage display
Phages
Presenilin 1
Receptors
Research and Analysis Methods
Screening
Senile plaques
Site-directed mutagenesis
Transferrin
Transferrins
Transgenic mice
Viral antibodies
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Summary:The blood-brain barrier (BBB) presents a major obstacle in developing specific diagnostic imaging agents for many neurological disorders. In this study we aimed to generate single domain anti-mouse transferrin receptor antibodies (anti-mTfR VHHs) to mediate BBB transcytosis as components of novel MRI molecular contrast imaging agents. Anti-mTfR VHHs were produced by immunizing a llama with mTfR, generation of a VHH phage display library, immunopanning, and in vitro characterization of candidates. Site directed mutagenesis was used to generate additional variants. VHH fusions with neurotensin (NT) allowed rapid, hypothermia-based screening for VHH-mediated BBB transcytosis in wild-type mice. One anti-mTfR VHH variant was fused with an anti-amyloid-beta (A[beta]) VHH dimer and labeled with fluorescent dye for direct assessment of in vivo target engagement in a mouse model of AD-related A[beta] plaque pathology. An anti-mTfR VHH called M1 and variants had binding affinities to mTfR of
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0276107