title: Efficient generation of a reshaped human mAb specific for the alpha toxin of Clostridium perfringens.
abstract: We have used the technique of antibody reshaping to produce a humanized antibody specific for the alpha toxin of Clostridium perfringens. The starting antibody was from a mouse hybridoma from which variable (V) region nucleotide sequences were determined. The complementarity-determining regions (CDRs) from these V regions were then inserted into human heavy and light chain V region genes with human constant region gene fragments subsequently added. The insertion of CDRs alone into human frameworks did not produce a functional reshaped antibody and modifications to the V region framework were required. With minor framework modifications, the affinity of the original murine mAb was restored and even exceeded. Where affinity was increased, an altered binding profile to overlapping peptides was observed. Computer modelling of the reshaped heavy chain V regions suggested that amino acids adjacent to CDRs can either contribute to, or distort, CDR loop conformation and must be adjusted to achieve high binding affinity.
authors.affiliation: Scotgen Biopharmaceuticals Inc., Aberdeen, UK.
authors.forename: P R
authors.initials: PR
authors.lastname: Tempest
authors.forename: P
authors.initials: P
authors.lastname: White
authors.forename: E D
authors.initials: ED
authors.lastname: Williamson
authors.forename: R W
authors.initials: RW
authors.lastname: Titball
authors.forename: D C
authors.initials: DC
authors.lastname: Kelly
authors.forename: G J
authors.initials: GJ
authors.lastname: Kemp
authors.forename: P M
authors.initials: PM
authors.lastname: Gray
authors.forename: S J
authors.initials: SJ
authors.lastname: Forster
authors.forename: F J
authors.initials: FJ
authors.lastname: Carr
authors.forename: W J
authors.initials: WJ
authors.lastname: Harris
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D001427:Bacterial Toxins / Q000737:chemistry* / Q000276:immunology*; D001483:Base Sequence; D001666:Binding Sites, Antibody; D002135:Calcium-Binding Proteins; D003016:Clostridium perfringens; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000737:chemistry*; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D016415:Sequence Alignment; D010738:Type C Phospholipases
chemical_list: D000911:Antibodies, Monoclonal; D001427:Bacterial Toxins; D002135:Calcium-Binding Proteins; D007135:Immunoglobulin Variable Region; D010455:Peptides; D011993:Recombinant Fusion Proteins; D010738:Type C Phospholipases; C040649:alpha toxin, Clostridium perfringens