Recombinant Human Antibody (S77) is capable of binding to C3b, expressed in HEK 293 cells. Expressed as the combination of a heavy chain (HC) containing VH from anti-C3b mAb and CH1-3 region of human IgG1 and a light chain (LC) encoding VL from anti-C3b proteins mAb and CL of human kappa light chain. Exists as a disulfide linked dimer of the HC and LC hetero-dimer under non-reducing condition. This antibody blocks binding of factor B to C3b inhibiting the first step in the formation of the alternative pathway C3 convertase.
Figure 1 Generation of a phage antibody that selectively binds to C3b, but not native C3.
Binding of S77 IgG to C3b. Binding is expressed as the ratio of optical densities of the reaction mixtures measured at 450 and 630 nm.
Katschke, K. J., Stawicki, S., Yin, J., Steffek, M., Xi, H., Sturgeon, L., ... & van Lookeren Campagne, M. (2009). Structural and functional analysis of a C3b-specific antibody that selectively inhibits the alternative pathway of complement. Journal of Biological Chemistry, 284(16), 10473-10479.
Figure 2 S77 inhibits fH and sCR1 binding to C3 and inhibits fH and sCR1co-factor activity.
Microtiter plates were coated with C3b, and fH wasadded in the presence of increasing concentrations of S77 or control Fab. Binding of fH to C3b was determined using an anti-fH antibody and a secondary HRPO-conjugated antibody. Absorbance of the reaction mixture was measured at 450 nm.
Katschke, K. J., Stawicki, S., Yin, J., Steffek, M., Xi, H., Sturgeon, L., ... & van Lookeren Campagne, M. (2009). Structural and functional analysis of a C3b-specific antibody that selectively inhibits the alternative pathway of complement. Journal of Biological Chemistry, 284(16), 10473-10479.
Figure 3 Cofactor activity for fI-mediated cleavage of C3b was measured by incubating C3b and fI with fH or increasing concentrations of S77 or control Fab.
The mixture was incubated at 37 °C, and the samples were analyzed by gel-electrophoresis and Simply Blue staining.
Katschke, K. J., Stawicki, S., Yin, J., Steffek, M., Xi, H., Sturgeon, L., ... & van Lookeren Campagne, M. (2009). Structural and functional analysis of a C3b-specific antibody that selectively inhibits the alternative pathway of complement. Journal of Biological Chemistry, 284(16), 10473-10479.
Figure 4 S77 inhibits the alternative, but not classical, pathway C5 convertase in human serum.
S77 does not inhibit CP convertase activation. IgM-coated sheep erythrocytes were incubated with factor B-depleted human serum and increasing concentrations of S77 or anti-C5 antibody.
Katschke, K. J., Stawicki, S., Yin, J., Steffek, M., Xi, H., Sturgeon, L., ... & van Lookeren Campagne, M. (2009). Structural and functional analysis of a C3b-specific antibody that selectively inhibits the alternative pathway of complement. Journal of Biological Chemistry, 284(16), 10473-10479.
Figure 5 S77 inhibits the alternative pathway C3 and C5 convertase.
C3 and factors B and D were incubated in the presence of increasing concentrations of S77 or control Fab. The concentration of C3a des-Arg reaction product was determined by ELISA.
Katschke, K. J., Stawicki, S., Yin, J., Steffek, M., Xi, H., Sturgeon, L., ... & van Lookeren Campagne, M. (2009). Structural and functional analysis of a C3b-specific antibody that selectively inhibits the alternative pathway of complement. Journal of Biological Chemistry, 284(16), 10473-10479.
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CAT | Product Name | Application | Type |
---|---|---|---|
PFBL-417 | Human Anti-C3b Recombinant Antibody (clone S77); Fab Fragment | ELISA | Human Fab |
HPAB-AP270-YC-F(E) | Human Anti-C3b Recombinant Antibody; Fab Fragment (HPAB-AP270-YC-F(E)) | ELISA, Inhib | Chimeric (mouse/human) Fab |
HPAB-AP271-YC-F(E) | Human Anti-C3b Recombinant Antibody; Fab Fragment (HPAB-AP271-YC-F(E)) | ELISA, Inhib | Humanized (mouse/human) Fab |
HPAB-AP272-YC-F(E) | Human Anti-C3b Recombinant Antibody; Fab Fragment (HPAB-AP272-YC-F(E)) | ELISA, Inhib | Humanized (mouse/human) Fab |
HPAB-0954WJ-F(E) | Human Anti-C3b Recombinant Antibody (clone MOR09556); Fab Fragment | ELISA | Human Fab |
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