Conformation-specific nanobody targeting alpha-synuclein oligomers for neurodegeneration theragnostics

Overview

This solution is a conformation-specific nanobody (an antibody fragment) designed to selectively target toxic oligomeric self-assemblies of the protein alpha-synuclein, which are implicated in neurodegeneration, including Parkinson's disease and Alzheimer's disease. Alpha-synuclein oligomers are small protein assemblies that trigger brain inflammation and damage neurons, making them key therapeutic targets and biomarkers. The nanobody binds to a specific region of alpha-synuclein called P1 (residues 36–42), which is exposed in oligomers but not in less toxic forms. This selective binding enables novel clinical theragnostic opportunities, combining therapeutic and diagnostic capabilities. The technology was developed by a team with a strong track record in integrative antibody discovery platforms targeting protein conformations and chemical modifications.

Technical specifications

• Selective binding: The nanobody specifically binds the P1 region (residues 36–42) of alpha-synuclein and does not cross-react with other proteins such as amyloid-beta.
• Conformation preference: The nanobody preferentially binds toxic oligomers rather than monomers or larger, less toxic assemblies of alpha-synuclein.
• Inhibition of self-assembly: Using FRET-based binding experiments and ELISA, the nanobody has been shown to inhibit alpha-synuclein self-assembly in buffer and under competition conditions (in the presence of cellular crude extracts) at sub-stoichiometric concentrations (1 nanobody per 100 alpha-synuclein molecules).
• Validation methods: Binding specificity confirmed by Förster resonance energy transfer (FRET) and Enzyme-Linked Immunosorbent Assay (ELISA); self-assembly inhibition confirmed using a fluorescent probe sensing protein self-assemblies and transmission electron microscopy (TEM).
• Discovery platform: Generated using an integrative antibody discovery platform capable of targeting protein conformations and chemical modifications.
• Intellectual property: A patent is being filed covering the nanobody, the discovery strategy, and in vitro binding characterization.

Technology readiness level

The nanobody has been validated in vitro with demonstrated specific binding to the P1 region and preferential binding to oligomers over monomers and larger assemblies. Self-assembly inhibition has been confirmed under both buffer and competition conditions. Future validation plans include using the nanobody to pull-down oligomers from in vitro aggregations and patient biofluids for screening-based small-molecule drug discovery (months 1–18), and testing therapeutic effects in neuroblastoma cells (SH-SY5Y) exposed to alpha-synuclein oligomers using membrane disruption and mitochondrial functionality assays (months 12–24). The technology is at an early-to-mid stage of preclinical development, with in vitro proof-of-concept established and cellular validation underway.