The Journal of Immunology · 1992 · 146 citations · 0 references
PharmacyImmunologyMolecular BiologyAntibody EngineeringMonoclonal AntibodiesProtein EngineeringAntibody CharacterizationAntibody DiscoveryAntibody StructureAntibody-antigen RecognitionAntigen DesignChimeric AntibodiesAmino AcidsAntibodiesAntibody Binding SpecificityAntigen-antibody RecognitionChimeric Monoclonal AntibodiesHumanized AntibodiesHumanized Monoclonal AntibodiesProtein Binding Affinity
L and H chain cDNAs of M195, a murine mAb that binds to the CD33 Ag on normal and leukemic myeloid cells, were cloned. The cDNAs were used in the construction of mouse/human IgG1 and IgG3 chimeric antibodies. In addition, humanized antibodies were constructed which combined the complementarity-determining regions of the M195 antibody with human framework and constant regions. The human framework was chosen to maximize homology with the M195 V domain sequence. Moreover, a computer model of M195 was used to identify several framework amino acids that are likely to interact with the complementarity-determining regions, and these residues were also retained in the humanized antibodies. Unexpectedly, the humanized IgG1 and IgG3 M195 antibodies, which have reshaped V regions, have higher apparent binding affinity for the CD33 Ag than the chimeric or mouse antibodies.