Blanco Toribio, AnaSainz Pastor, NoeliaÁlvarez Cienfuegos, AnaMerino, NekaneSánchez Martín, DavidBonet, JaumeSantos Valle, PatriciaSanz, LauraOliva, BaldoBlanco, Francisco J.Álvarez Vallina, LuisCuesta Martínez, Ángel2024-01-182024-01-182013Blanco-Toribio A, Sainz-Pastor N, Álvarez-Cienfuegos A, et al. Generation and characterization of monospecific and bispecific hexavalent trimerbodies. MAbs. 2013;5(1):70-79. doi:10.4161/mabs.226981942-086210.4161/mabs.22698https://hdl.handle.net/20.500.14352/93800Here, we describe a new class of multivalent and multispecific antibody-based reagents for therapy. The molecules, termed “trimerbodies,” use a modified version of the N-terminal trimerization region of human collagen XVIII noncollagenous 1 domain flanked by two flexible linkers as trimerizing scaffold. By fusing single-chain variable fragments (scFv) with the same or different specificity to both N- and C-terminus of the trimerizing scaffold domain, we produced monospecific or bispecific hexavalent molecules that were efficiently secreted as soluble proteins by transfected mammalian cells. A bispecific anti-laminin x anti-CD3 N-/C-trimerbody was found to be trimeric in solution, very efficient at recognizing purified plastic-immobilized laminin and CD3 expressed at the surface of T cells, and remarkably stable in human serum. The bispecificity was further demonstrated in T cell activation studies. In the presence of laminin-rich substrate, the bispecific anti-laminin x anti-CD3 N-/C-trimerbody stimulated a high percentage of human T cells to express surface activation markers. These results suggest that the trimerbody platform offers promising opportunities for the development of the next-generation therapeutic antibodies, i.e., multivalent and bispecific molecules with a format optimized for the desired pharmacokinetics and adapted to the pathological context.engAttribution-NonCommercial 4.0 Internationalhttp://creativecommons.org/licenses/by-nc/4.0/Generation and characterization of monospecific and bispecific hexavalent trimerbodiesjournal article1942-0870https://doi.org/10.4161/mabs.22698https://www.tandfonline.com/doi/abs/10.4161/mabs.22698open access577.1577.2Antibody engineeringTrimerbodyCollagenBispecific antibodiesMultivalent antibodiesBiología molecular (Farmacia)Bioquímica (Farmacia)24 Ciencias de la Vida