title: Genetically engineered deglycosylation of the variable domain increases the affinity of an anti-CD33 monoclonal antibody.
abstract: M195 is a murine monoclonal antibody that binds to the CD33 antigen and is being tested for the treatment of myeloid leukemia. Surprisingly, a complementarity determining region (CDR)-grafted, humanized M195 antibody displayed a several-fold higher binding affinity for the CD33 antigen than the original murine antibody. Here we show that the increase in binding affinity resulted from eliminating an N-linked glycosylation site at residue 73 in the heavy chain variable region in the course of humanization. Re-introducing the glycosylation site in the humanized antibody reduces its binding affinity to that of the murine antibody, while removing the glycosylation site from the murine M195 variable domain increases its affinity. The removal of variable region carbohydrates may provide a method for increasing the affinity of certain monoclonal antibodies with diagnostic and therapeutic potential.
authors.affiliation: Protein Design Labs, Mountain View, CA 94043.
authors.forename: M S
authors.initials: MS
authors.lastname: Co
authors.forename: D A
authors.initials: DA
authors.lastname: Scheinberg
authors.forename: N M
authors.initials: NM
authors.lastname: Avdalovic
authors.forename: K
authors.initials: K
authors.lastname: McGraw
authors.forename: M
authors.initials: M
authors.lastname: Vasquez
authors.forename: P C
authors.initials: PC
authors.lastname: Caron
authors.forename: C
authors.initials: C
authors.lastname: Queen
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D000915:Antibody Affinity; D015703:Antigens, CD / Q000276:immunology*; D015214:Antigens, Differentiation, Myelomonocytic / Q000276:immunology*; D001666:Binding Sites, Antibody; D006023:Glycoproteins / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000737:chemistry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000276:immunology; D063268:Sialic Acid Binding Ig-like Lectin 3; D013329:Structure-Activity Relationship
chemical_list: D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D015214:Antigens, Differentiation, Myelomonocytic; D006023:Glycoproteins; D007135:Immunoglobulin Variable Region; D011993:Recombinant Fusion Proteins; D063268:Sialic Acid Binding Ig-like Lectin 3