title: Crystal structure of an anti-Ang2 CrossFab demonstrates complete structural and functional integrity of the variable domain.
abstract: Bispecific antibodies are considered as a promising class of future biotherapeutic molecules. They comprise binding specificities for two different antigens, which may provide additive or synergistic modes of action. There is a wide variety of design alternatives for such bispecific antibodies, including the "CrossMab" format. CrossMabs contain a domain crossover in one of the antigen-binding (Fab) parts, together with the "knobs-and-holes" approach, to enforce the correct assembly of four different polypeptide chains into an IgG-like bispecific antibody. We determined the crystal structure of a hAng-2-binding Fab in its crossed and uncrossed form and show that CH1-CL-domain crossover does not induce significant perturbations of the structure and has no detectable influence on target binding.
authors.affiliation: Large Molecule Research, Pharma Research and Early Development, Roche Dignostics GmbH, Penzberg, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Fenn
authors.forename: Christian B
authors.initials: CB
authors.lastname: Schiller
authors.forename: Julia J
authors.initials: JJ
authors.lastname: Griese
authors.forename: Harald
authors.initials: H
authors.lastname: Duerr
authors.forename: Sabine
authors.initials: S
authors.lastname: Imhof-Jung
authors.forename: Christian
authors.initials: C
authors.lastname: Gassner
authors.forename: Joerg
authors.initials: J
authors.lastname: Moelleken
authors.forename: Joerg Thomas
authors.initials: JT
authors.lastname: Regula
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Schaefer
authors.forename: Markus
authors.initials: M
authors.lastname: Thomas
authors.forename: Christian
authors.initials: C
authors.lastname: Klein
authors.forename: Karl-Peter
authors.initials: KP
authors.lastname: Hopfner
authors.forename: Hubert
authors.initials: H
authors.lastname: Kettenberger
mesh_terms: D000595:Amino Acid Sequence; D042702:Angiopoietin-2 / Q000276:immunology*; D018033:Antibodies, Bispecific / Q000737:chemistry* / Q000378:metabolism*; D018360:Crystallography, X-Ray; D057809:HEK293 Cells; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry* / Q000378:metabolism*; D007135:Immunoglobulin Variable Region / Q000737:chemistry* / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D055550:Protein Stability; D017434:Protein Structure, Tertiary; D055672:Static Electricity; D013329:Structure-Activity Relationship; D013696:Temperature
chemical_list: D042702:Angiopoietin-2; D018033:Antibodies, Bispecific; D007140:Immunoglobulin Fab Fragments; D007135:Immunoglobulin Variable Region