title: Engineering a therapeutic IgG molecule to address cysteinylation, aggregation and enhance thermal stability and expression.
abstract: Antibodies can undergo a variety of covalent and non-covalent degradation reactions that have adverse effects on efficacy, safety, manufacture and storage. We had identified an antibody to Angiopoietin 2 (Ang2 mAb) that neutralizes Ang2 binding to its receptor in vitro and inhibits tumor growth in vivo. Despite favorable pharmacological activity, the Ang2 mAb preparations were heterogeneous, aggregated rapidly and were poorly expressed. Here, we report the engineering of the antibody variable and constant domains to generate an antibody with reduced propensity to aggregate, enhanced homogeneity, 11°C elevated T(m), 26-fold improved level of expression and retained activity. The engineered molecule, MEDI-3617, is now compatible with the large scale material supply required for clinical trials and is currently being evaluated in Phase 1 in cancer patients. This is the first report to describe the stability engineering of a therapeutic antibody addressing non canonical cysteine residues and the design strategy reported here is generally applicable to other therapeutic antibodies and proteins.
authors: MedImmune, Cambridge, UK. buchanana@medimmune.com
authors: Andrew
authors: A
authors: Buchanan
authors: Veronica
authors: V
authors: Clementel
authors: Rob
authors: R
authors: Woods
authors: Nicholas
authors: N
authors: Harn
authors: Michael A
authors: MA
authors: Bowen
authors: Wenjun
authors: W
authors: Mo
authors: Bojana
authors: B
authors: Popovic
authors: Steven M
authors: SM
authors: Bishop
authors: William
authors: W
authors: Dall'Acqua
authors: Ralph
authors: R
authors: Minter
authors: Lutz
authors: L
authors: Jermutus
authors: Vahe
authors: V
authors: Bedian
mesh_terms: D042702:Angiopoietin-2 / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism / Q000627:therapeutic use; D003545:Cysteine / Q000378:metabolism; D006358:Hot Temperature; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism* / Q000627:therapeutic use; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D015202:Protein Engineering / Q000379:methods*; D055550:Protein Stability
chemical_list: D042702:Angiopoietin-2; D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D003545:Cysteine
keywords: aggregation; antibody; cysteinylation; expression; stability