Background
Assembling nanobodies (Nb) into polyvalent multimers is a powerful strategy for improving the effectiveness of Nb-based therapeutics and biotechnological tools. However, generally effective approaches to Nb assembly are currently restricted to the N- or C-termini, greatly limiting the diversity of Nb multimers that can be produced.
Here, Oregon State University researchers show that reactive tetrazine groups – site-specifically inserted by genetic code expansion (GCE) at Nb surface sites – are compatible with Nb folding and function.
This tetrazine-based approach is a generally applicable strategy that greatly increases the diversity of accessible Nb-assemblies and adds assembly geometry as something that can be easily varied to optimize the properties of Nb-assemblies.
Technology Description
A Multimeric nanobody assembly made up of at least two or more tetrazine functionalized nanobodies (two and three multivalent options are shown to work) connected by a non peptide linker. The GCE-Tetrazine encoding technology provides both optimal chemistry and unfettered attachment points to functionalize Nbs with no restrictions from bioorthogonal conjugation conditions. The conjugation approach allows researchers to access the full combinatorial diversity of Nb assemblies with facile modulation of the sites of conjugation on Nbs, the linker lengths and geometries and the types of Nb used.
Further Details: Nanobody assemblies with fully flexible topology enabled by genetically encoded tetrazine amino acids
Science Advances, 6 May 2022, Vol 8, Issue 18
Features & Benefits
This rapid assembly and screening of combinatorial fused Nb architectures will advantage therapeutic Nb development when optimizing therapeutic window, lifetime, toxicity, etc. and will provide a unique advantage when combating rapidly evolving viral targets, such as SARS-CoV-2.
Applications
- Therapeutics
- Research Tools
Status
U.S. non-provisional patent application No. 18/061,389