INHIBITION OF COMPLEMENT ACTIVITY BY HUMANIZED ANTI-C5 ANTIBODY AND SINGLE-CHAIN FV

Activation of the complement system contributes significantly to the p athogenesis of numerous acute and chronic diseases. Recently, a monocl onal antibody (5G1.1) that recognizes the human complement protein C5, has been shown to effectively block C5 cleavage, thereby preventing t he generation of the pro-inflammatory complement components C5a and C5 b-9. Humanized 5G1.1 antibody, Fab and scFv molecules have been produc ed by grafting the complementarity determining regions of 5G1.1 on to human framework regions. Competitive ELISA analysis indicated that no framework changes were required in the humanized variable regions for retention of high affinity binding to C5, even at framework positions predicted by computer modeling to influence CDR canonical structure. T he humanized Fab and scFv molecules blocked complement-mediated lysis of chicken erythrocytes and porcine aortic endothelial cells in a dose -dependent fashion, with complete complement inhibition occurring at a three-fold molar excess, relative to the human C5 concentration. In c ontrast to a previously characterized anti-C5 scFv molecule, the human ized h5G1.1 scFv also effectively blocked C5a generation. Finally, an intact humanized h5G1.1 antibody blocked human complement lytic activi ty at concentrations identical to the original murine monoclonal antib ody. These results demonstrate that humanized h5G1.1 and its recombina nt derivatives retain both the affinity and blocking functions of the murine 5G1.1 antibody, and suggest that these molecules may serve as p otent inhibitors of complement-mediated pathology in human inflammator y diseases. (C) 1997 Elsevier Science Ltd.