%0 Journal Article %1 Fellermeier2016 %A Fellermeier, Sina %A Beha, Nadine %A Meyer, Jan Erik %A Ring, Sarah %A Bader, Stefan %A Kontermann, Roland E. %A Müller, Dafne %D 2016 %I Taylor & Francis %J OncoImmunology %K 2016 izi kontermann %N 11 %P e1238540 %R 10.1080/2162402X.2016.1238540 %T Advancing targeted co-stimulation with antibody-fusion proteins by introducing TNF superfamily members in a single-chain format %U https://www.tandfonline.com/doi/full/10.1080/2162402X.2016.1238540 %V 5 %X © 2016 Taylor & Francis Group, LLCCo-stimulation via receptors of the tumor necrosis factor superfamily (TNFSF) emerges as promising strategy to support antitumor immune responses. Targeted strategies with antibody-fusion proteins composed of a tumor-directed antibody part and the extracellular domain of a co-stimulatory ligand of the TNFSF constitute an attractive option to focus the co-stimulatory activity to the tumor site. Since TNFSF members intrinsically form functional units of non-covalently linked homotrimers, the protein engineering of suitable antibody-fusion proteins is challenging. Aiming for molecules of simple and stable configuration, we used TNFSF ligands in a single-chain format (scTNFSF), i.e., three units of the ectodomain connected by polypeptide linkers, folding into an intramolecular trimer. By fusing tumor-directed scFv antibody fragments directed against EpCAM or FAP to co-stimulatory scTNFSF molecules (sc4-1BBL, scOX40L, scGITRL or scLIGHT), a set of monomeric scFv-scTNFSF fusion proteins was generated. In comparison to the scFv-TNFSF format, defined by intermolecular homotrimerization via the TNFSF part, scFv-scTNFSF showed equal or enhanced co-stimulatory activity despite reduced avidity in antibody binding. In addition, enhanced serum stability and improved bioavailability in mice were observed. We show that the scFv-scTNFSF format can be applied to various members of the TNFSF, presenting targeting-dependent co-stimulatory activity. Hence, this format exhibits favorable properties that make it a promising choice for further therapeutic fusion protein development.

@article{Fellermeier2016, abstract = {{\textcopyright} 2016 Taylor {\&} Francis Group, LLCCo-stimulation via receptors of the tumor necrosis factor superfamily (TNFSF) emerges as promising strategy to support antitumor immune responses. Targeted strategies with antibody-fusion proteins composed of a tumor-directed antibody part and the extracellular domain of a co-stimulatory ligand of the TNFSF constitute an attractive option to focus the co-stimulatory activity to the tumor site. Since TNFSF members intrinsically form functional units of non-covalently linked homotrimers, the protein engineering of suitable antibody-fusion proteins is challenging. Aiming for molecules of simple and stable configuration, we used TNFSF ligands in a single-chain format (scTNFSF), i.e., three units of the ectodomain connected by polypeptide linkers, folding into an intramolecular trimer. By fusing tumor-directed scFv antibody fragments directed against EpCAM or FAP to co-stimulatory scTNFSF molecules (sc4-1BBL, scOX40L, scGITRL or scLIGHT), a set of monomeric scFv-scTNFSF fusion proteins was generated. In comparison to the scFv-TNFSF format, defined by intermolecular homotrimerization via the TNFSF part, scFv-scTNFSF showed equal or enhanced co-stimulatory activity despite reduced avidity in antibody binding. In addition, enhanced serum stability and improved bioavailability in mice were observed. We show that the scFv-scTNFSF format can be applied to various members of the TNFSF, presenting targeting-dependent co-stimulatory activity. Hence, this format exhibits favorable properties that make it a promising choice for further therapeutic fusion protein development.}, added-at = {2023-06-29T13:07:55.000+0200}, author = {Fellermeier, Sina and Beha, Nadine and Meyer, Jan Erik and Ring, Sarah and Bader, Stefan and Kontermann, Roland E. and M{\"{u}}ller, Dafne}, biburl = {https://puma.ub.uni-stuttgart.de/bibtex/290b0319e32a9e0ba019cc416dd1ce10f/fabian}, doi = {10.1080/2162402X.2016.1238540}, interhash = {dfbb566cca0bc7a52c0fed97637ad90c}, intrahash = {90b0319e32a9e0ba019cc416dd1ce10f}, issn = {2162402X}, journal = {OncoImmunology}, keywords = {2016 izi kontermann}, month = nov, number = 11, pages = {e1238540}, pmid = {27999756}, publisher = {Taylor {\&} Francis}, timestamp = {2023-06-29T13:07:55.000+0200}, title = {{Advancing targeted co-stimulation with antibody-fusion proteins by introducing TNF superfamily members in a single-chain format}}, url = {https://www.tandfonline.com/doi/full/10.1080/2162402X.2016.1238540}, volume = 5, year = 2016 }