A humanized antibody specific for the platelet integrin gpIIb/IIIa.
C4G1, a murine mAb reactive with the platelet gpIIb/IIIa integrin, was humanized for potential treatment of thrombosis-related disorders. The variable regions of light- and heavy-chain cDNAs from the C4G1 hybridoma were first cloned and sequenced. Humanized C4G1 Ab of the IgG1 isotype was constructed by combining the complementarity-determining regions of C4G1 with human framework and constant regions. The human framework was chosen to maximize homology with the C4G1 variable region sequence, and a computer model of C4G1 was used to aid design of the final framework sequence. Genetic constructs were also developed to produce Fab and F(ab')2 fragments of the humanized C4G1 Ab. The humanized IgG1 Ab as well as the Fab and F(ab')2 fragments showed equivalent binding affinities to their murine counterparts, indicating no loss in binding affinity during the humanization process. The humanized Ab and its fragments were also shown to inhibit platelet aggregation and to inhibit binding of fibrinogen to gpIIb/IIIa in vitro.
Co
M S
MS
Protein Design Labs, Inc., Mountain View, CA 94043.
Yano
S
S
Hsu
R K
RK
Landolfi
N F
NF
Vasquez
M
M
Cole
M
M
Tso
J T
JT
Bringman
T
T
Laird
W
W
Hudson
D
D
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D003001:Cloning, Molecular; D003198:Computer Simulation; D005340:Fibrinogen / Q000378:metabolism; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis; D051379:Mice; D008969:Molecular Sequence Data; D010974:Platelet Aggregation; D010980:Platelet Membrane Glycoproteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D010980:Platelet Membrane Glycoproteins; D005340:Fibrinogen