Nanobodies raised against the cytotoxic α-synuclein oligomer are oligomer-specific and promote its cellular uptake
Parkinson’s disease involves the accumulation of aggregates of ɑ-synuclein (ɑ-Syn), both as intracellular fibrils and as cytotoxic soluble oligomeric species (ɑSOs). No available nanobodies show exclusive preference for the oligomeric state of ɑ-Syn. Here, we describe two nanobodies NB1 and NB2, obtained by immunizing a llama with αSOs, which bind ɑSOs with nM affinity and do not show any measurable affinity for monomeric ɑ-Syn or ɑ-Syn fibrils. While the nanobodies were not useful for high-throughput screening for therapeutic compounds or high-resolution cryoEM, they retained their ability to discriminate against ɑ-Syn monomers in brain tissue and were able to detect ɑ-Syn aggregates in diseased tissue. In addition, αSO binding affinity was improved by DNA-scaffold-mediated NB1 dimerization compared to scaffolded monomeric NB1. The nanobodies promote the uptake of ɑSOs into HEK93 cells via the Sortilin receptor pathway. Their absolute specificity for oligomeric ɑ-Syn makes them promising reagents to detect oligomeric ɑ-Syn in patient samples.
Authors
- Camilla Gustafsen (ORCID: https://orcid.org/0000-0002-3349-4639)
- Mette Galsgaard Malle (ORCID: https://orcid.org/0000-0003-3722-502X)
- Charlie Johansen
- Marianne Kristensen
- Jannik Nedergaard Pedersen (ORCID: https://orcid.org/0000-0003-1416-5626)
- Brian C. Berg
- Marina Romero‐Ramos (ORCID: https://orcid.org/0000-0003-0970-578X)
- Kasper R. Andersen (ORCID: https://orcid.org/0000-0002-4415-8067)
- William P. Olsen (ORCID: https://orcid.org/0000-0002-5603-8310)
- Elisa Sassetti (ORCID: https://orcid.org/0000-0001-9816-5636)
- Peder Madsen (ORCID: https://orcid.org/0000-0002-8845-4802)
- Julián Valero (ORCID: https://orcid.org/0000-0002-5260-2770)
- Brandi Simonsen (ORCID: https://orcid.org/0000-0002-1561-1686)
- Johanne Lauritsen
- Andreas Bøggild (ORCID: https://orcid.org/0000-0003-2130-3521)
- Kristian Strømgaard (ORCID: https://orcid.org/0000-0003-2206-4737)
- Amanda Simonsen
- Jakob Hauge Mikkelsen (ORCID: https://orcid.org/0000-0002-2914-3297)
- Thomas Boesen (ORCID: https://orcid.org/0000-0002-5633-6844)
- Mads H. Clausen (ORCID: https://orcid.org/0000-0001-9649-1729)
- Simon Glerup (ORCID: https://orcid.org/0000-0002-9686-8124)
- Jørgen Kjems (ORCID: https://orcid.org/0000-0003-4128-9317)
- Svend Birkelund (ORCID: https://orcid.org/0000-0001-6171-5606)
- Janni Nielsen
- J. Carlson (ORCID: https://orcid.org/0000-0002-3163-5565)
- Daniel E. Otzen (ORCID: https://orcid.org/0000-0002-2918-8989)
- Simon B. Hansen (ORCID: https://orcid.org/0000-0002-0843-3922)
- Giulia Kleijwegt
- Faranak Nami (ORCID: https://orcid.org/0000-0002-4152-1318)
- Gunna Christiansen (ORCID: https://orcid.org/0000-0003-0540-1367)
- Jan Stanislaw Nowak (ORCID: https://orcid.org/0009-0000-8260-4622)
- Bo Volf Brøchner
- Vita Sereika-Bejder
- Kirstine Friis Merrild
- Nanna Miang Lyngsø
Institutions
- University of Copenhagen (DK)
- Aarhus University (DK)
- Aarhus University Hospital (DK)
- Aalborg University (DK)
- Technical University of Denmark (DK)
Publication Details
- Journal
- npj Biosensing
- Published
- 2025-06-02
- DOI
- https://doi.org/10.1038/s44328-025-00042-1
- Citations
- 2
- Primary Topic
- Monoclonal and Polyclonal Antibodies Research
- Type
- article
- Field-Weighted Citation Impact
- 2.72
Funders
- Aarhus Universitet
- Uddannelses- og Forskningsministeriet
- National Research Foundation
- Danmarks Grundforskningsfond
- Carlsbergfondet
- Lundbeckfonden
- Novo Nordisk
- Danmarks Frie Forskningsfond
- Danmarks Tekniske Universitet
- Novo Nordisk Fonden
- Danmarks Frie Forskningsfond
- H. Lundbeck A/S