%0 Journal Article %1 Stork2008 %A Stork, Roland %A Zettlitz, Kirstin A. %A Müller, Dafne %A Rether, Miriam %A Hanisch, Franz Georg %A Kontermann, Roland E. %D 2008 %J Journal of Biological Chemistry %K 2008 izi kontermann %N 12 %P 7804--7812 %R 10.1074/jbc.M709179200 %T N-glycosylation as novel strategy to improve pharmacokinetic properties of bispecific single-chain diabodies %U https://www.ncbi.nlm.nih.gov/pubmed/18211902 %V 283 %X The therapeutic efficacy of recombinant antibodies such as single-chain Fv fragments and small bispecific or bifunctional molecules is often limited by rapid elimination from the circulation because of their small size. Here, we have investigated the effects of N-glycosylation on the activity and pharmacokinetics of a small bispecific single-chain diabody (scDb CEACD3) developed for the retargeting of cytotoxic T cells to CEA-expressing tumor cells. We could show that the introduction of N-glycosylation sequons into the flanking linker and a C-terminal extension results in the production of N-glycosylated molecules after expression in transfected HEK293 cells. N-Glycosylated scDb variants possessing 3, 6, or 9 N-glycosylation sites, respectively, retained antigen binding activity and bispecificity for target and effector cells as shown in a target cell-dependent IL-2 release assay, although activity was reduced approximately 3-5-fold compared with the unmodified scDb. All N-glycosylated scDb variants exhibited a prolonged circulation time compared with scDb, leading to a 2-3-fold increase of the area under curve (AUC). In comparison, conjugation of a branched 40-kDa PEG chain increased AUC by a factor of 10.6, while a chimeric anti-CEA IgG1 molecule had the longest circulation time with a 17-fold increase in AUC. Thus, N-glycosylation complements the repertoire of strategies to modulate pharmacokinetics of small recombinant antibody molecules by an approach that moderately prolongs circulation time. %Z Stork, RolandZettlitz, Kirstin AMuller, DafneRether, MiriamHanisch, Franz-GeorgKontermann, Roland EengResearch Support, Non-U.S. Gov'tJ Biol Chem. 2008 Mar 21;283(12):7804-12. doi: 10.1074/jbc.M709179200. Epub 2008 Jan 22. %7 2008/01/24 %@ 4971168567

@article{Stork2008, abstract = {The therapeutic efficacy of recombinant antibodies such as single-chain Fv fragments and small bispecific or bifunctional molecules is often limited by rapid elimination from the circulation because of their small size. Here, we have investigated the effects of N-glycosylation on the activity and pharmacokinetics of a small bispecific single-chain diabody (scDb CEACD3) developed for the retargeting of cytotoxic T cells to CEA-expressing tumor cells. We could show that the introduction of N-glycosylation sequons into the flanking linker and a C-terminal extension results in the production of N-glycosylated molecules after expression in transfected HEK293 cells. N-Glycosylated scDb variants possessing 3, 6, or 9 N-glycosylation sites, respectively, retained antigen binding activity and bispecificity for target and effector cells as shown in a target cell-dependent IL-2 release assay, although activity was reduced approximately 3-5-fold compared with the unmodified scDb. All N-glycosylated scDb variants exhibited a prolonged circulation time compared with scDb, leading to a 2-3-fold increase of the area under curve (AUC). In comparison, conjugation of a branched 40-kDa PEG chain increased AUC by a factor of 10.6, while a chimeric anti-CEA IgG1 molecule had the longest circulation time with a 17-fold increase in AUC. Thus, N-glycosylation complements the repertoire of strategies to modulate pharmacokinetics of small recombinant antibody molecules by an approach that moderately prolongs circulation time.}, added-at = {2023-06-29T13:07:55.000+0200}, annote = {Stork, RolandZettlitz, Kirstin AMuller, DafneRether, MiriamHanisch, Franz-GeorgKontermann, Roland EengResearch Support, Non-U.S. Gov'tJ Biol Chem. 2008 Mar 21;283(12):7804-12. doi: 10.1074/jbc.M709179200. Epub 2008 Jan 22.}, author = {Stork, Roland and Zettlitz, Kirstin A. and M{\"{u}}ller, Dafne and Rether, Miriam and Hanisch, Franz Georg and Kontermann, Roland E.}, biburl = {https://puma.ub.uni-stuttgart.de/bibtex/2d75239a53149873995fdeebcab822e0c/fabian}, doi = {10.1074/jbc.M709179200}, edition = {2008/01/24}, interhash = {2d8e2a5eff6769723d0fd0abae468c74}, intrahash = {d75239a53149873995fdeebcab822e0c}, isbn = {4971168567}, issn = {00219258}, journal = {Journal of Biological Chemistry}, keywords = {2008 izi kontermann}, number = 12, pages = {7804--7812}, pmid = {18211902}, timestamp = {2023-06-29T13:07:55.000+0200}, title = {{N-glycosylation as novel strategy to improve pharmacokinetic properties of bispecific single-chain diabodies}}, url = {https://www.ncbi.nlm.nih.gov/pubmed/18211902}, volume = 283, year = 2008 }