title: A single amino acid substitution abolishes the heterogeneity of chimeric mouse/human (IgG4) antibody.
abstract: Human immunoglobulin G4 (IgG4) exists in two molecular forms due to the heterogeneity of the inter-heavy chain disulphide bridges in the hinge region in a proportion of secreted human IgG4. This heterogeneity is only revealed under denaturing, non-reducing conditions in which an HL "half antibody" is detected, a phenomenon not seen in other human IgG isotypes. In native conditions noncovalent interactions hold the antibody together as the H2L2 tetramer. Analysis of the hinge sequences of human IgG heavy chains suggested that the presence of serine at residue 241 might be the cause of this heterogeneity. We therefore changed the serine at 241 to proline (found at that position in IgG1 and IgG2) in a mouse/human chimeric heavy chain. This single residue substitution leads to the production of a homogeneous antibody. Further, the variant IgG4 has significantly extended serum half-life and shows an improved tissue distribution compared to the original chimeric IgG4.
authors.affiliation: Celltech Ltd., Slough, Berks, U.K.
authors.forename: S
authors.initials: S
authors.lastname: Angal
authors.forename: D J
authors.initials: DJ
authors.lastname: King
authors.forename: M W
authors.initials: MW
authors.lastname: Bodmer
authors.forename: A
authors.initials: A
authors.lastname: Turner
authors.forename: A D
authors.initials: AD
authors.lastname: Lawson
authors.forename: G
authors.initials: G
authors.lastname: Roberts
authors.forename: B
authors.initials: B
authors.lastname: Pedley
authors.forename: J R
authors.initials: JR
authors.lastname: Adair
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000915:Antibody Affinity; D001667:Binding, Competitive; D002460:Cell Line; D006224:Cricetinae / Q000235:genetics; D004306:Dose-Response Relationship, Immunologic; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007074:Immunoglobulin G / Q000235:genetics* / Q000276:immunology*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000276:immunology; D007132:Immunoglobulin Isotypes / Q000235:genetics* / Q000276:immunology*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017354:Point Mutation; D011993:Recombinant Fusion Proteins; D017386:Sequence Homology, Amino Acid; D014162:Transfection
chemical_list: D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007132:Immunoglobulin Isotypes; D011993:Recombinant Fusion Proteins