Valence and avidity determine the agonistic activity of anti-TNFR2 nanobody fusion proteins
Year: 2025
Type: article
Abstract: We identified several TNFR2-specific nanobodies (Nbs). When formatted as dimeric Fc fusion proteins, these nanobodies exhibited no agonistic activity. To improve activity, we genetically fused one, two, or three copies of the TNFR2-specific Nb:188 to the constant regions of an IgG1 antibody, an Fab fragment, an Fc domain or the trimerization domain of tenascin-C resulting in constructs with 2-12 Nb:188 domains. Constructs with 2 or 3 Nb:188 domains displayed no or minimal activity, while those with 4 and 5 Nb:188 domains demonstrated moderate activity. However, constructs with 6 or more Nb:188 domains exhibited potent agonism, reaching half-maximal TNFR2 activation at concentrations in the low picomolar range. Similarly, hexameric constructs generated with other TNFR2-specific Nb domains demonstrated robust agonism, too. Benchmarking against various other ligand- and antibody-based TNFR2 agonists revealed that the hexameric 3xNb:188-Fc format displays superior specific activity and efficiently expands regulatory T cells (Tregs).
Source: Cell chemical biology
Authors: Mohamed A. Anany, Daniela Siegmund, Olena Zaitseva, Isabell Lang, Bayan Mouhandes, Mervat Salah, Daniela Weisenberger, Svetlana Stepanzow, Juan Gamboa Vargas, Julia Dahlhoff, Theresa Schneider, Giel Tanghe, Frederik Stevenaert, Tugsan Tezil, Tom Van Belle, Bipasa Kar, Thomas Dandekar, Heike Margarete Hermanns, Luc van Rompaey, Andreas Beilhack, Harald Wajant
Language: English
Subfield: Radiology, Nuclear Medicine and Imaging
Field: Medicine
Domain: Health Sciences
Open Access status: hybrid
Estimated APC paid: $9,350
Funder: Deutsche Forschungsgemeinschaft
Award: G6566832675
OpenAlex ID: W4414751303