title: Insertion of scFv into the hinge domain of full-length IgG1 monoclonal antibody results in tetravalent bispecific molecule with robust properties.
abstract: By simultaneous binding two disease mediators, bispecific antibodies offer the opportunity to broaden the utility of antibody-based therapies. Herein, we describe the design and characterization of Bs4Ab, an innovative and generic bispecific tetravalent antibody platform. The Bs4Ab format comprises a full-length IgG1 monoclonal antibody with a scFv inserted into the hinge domain. The Bs4Ab design demonstrates robust manufacturability as evidenced by MEDI3902, which is currently in clinical development. To further demonstrate the applicability of the Bs4Ab technology, we describe the molecular engineering, biochemical, biophysical, and in vivo characterization of a bispecific tetravalent Bs4Ab that, by simultaneously binding vascular endothelial growth factor and angiopoietin-2, inhibits their function. We also demonstrate that the Bs4Ab platform allows Fc-engineering similar to that achieved with IgG1 antibodies, such as mutations to extend half-life or modulate effector functions.
authors.affiliation: a Antibody Discovery and Protein Engineering , Gaithersburg , MA , USA.
authors.forename: Binyam
authors.initials: B
authors.lastname: Bezabeh
authors.affiliation: a Antibody Discovery and Protein Engineering , Gaithersburg , MA , USA.
authors.forename: Ryan
authors.initials: R
authors.lastname: Fleming
authors.affiliation: b Oncology Research , Gaithersburg , MA , USA.
authors.forename: Christine
authors.initials: C
authors.lastname: Fazenbaker
authors.affiliation: b Oncology Research , Gaithersburg , MA , USA.
authors.forename: Haihong
authors.initials: H
authors.lastname: Zhong
authors.affiliation: c Clinical Pharmacology and DMPK , Gaithersburg , MA , USA.
authors.forename: Karen
authors.initials: K
authors.lastname: Coffman
authors.affiliation: c Clinical Pharmacology and DMPK , Gaithersburg , MA , USA.
authors.forename: Xiang-Qing
authors.initials: XQ
authors.lastname: Yu
authors.affiliation: c Clinical Pharmacology and DMPK , Gaithersburg , MA , USA.
authors.forename: Ching Ching
authors.initials: CC
authors.lastname: Leow
authors.affiliation: a Antibody Discovery and Protein Engineering , Gaithersburg , MA , USA.
authors.forename: Nerea
authors.initials: N
authors.lastname: Gibson
authors.affiliation: a Antibody Discovery and Protein Engineering , Gaithersburg , MA , USA.
authors.forename: Susan
authors.initials: S
authors.lastname: Wilson
authors.affiliation: d Infectious Diseases, MedImmune , Gaithersburg , MA , USA.
authors.forename: C Kendall
authors.initials: CK
authors.lastname: Stover
authors.affiliation: a Antibody Discovery and Protein Engineering , Gaithersburg , MA , USA.
authors.forename: Herren
authors.initials: H
authors.lastname: Wu
authors.affiliation: a Antibody Discovery and Protein Engineering , Gaithersburg , MA , USA.
authors.forename: Changshou
authors.initials: C
authors.lastname: Gao
authors.affiliation: a Antibody Discovery and Protein Engineering , Gaithersburg , MA , USA.
authors.forename: Nazzareno
authors.initials: N
authors.lastname: Dimasi
mesh_terms: D042702:Angiopoietin-2 / Q000037:antagonists & inhibitors; D000818:Animals; D018033:Antibodies, Bispecific / Q000096:biosynthesis / Q000494:pharmacology*; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000494:pharmacology; D015179:Colorectal Neoplasms / Q000188:drug therapy; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000494:pharmacology*; D051379:Mice; D015202:Protein Engineering / Q000379:methods*; D057127:Single-Chain Antibodies / Q000096:biosynthesis / Q000494:pharmacology*; D042461:Vascular Endothelial Growth Factor A / Q000037:antagonists & inhibitors; D023041:Xenograft Model Antitumor Assays
chemical_list: C586513:ANGPT2 protein, human; D042702:Angiopoietin-2; D018033:Antibodies, Bispecific; D000911:Antibodies, Monoclonal; D000970:Antineoplastic Agents; D007074:Immunoglobulin G; D057127:Single-Chain Antibodies; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A
keywords: Antibody engineering; bispecific antibody; concurrent binding to antigens; multispecific antibody; single-chain antibody Fv; tetravalent antibody