This antibody binds to a unique conformational epitope in the native form of GPC3 core protein on cancer cells with high affinity. The binding requires both the N-terminus and C-terminus domains of GPC3, and is independent of the HS chains on GPC3.
Figure 1 Binding properties of HN3 antibody.
Protein-binding affinity was measured by ELISA using purified GPC3-hFc. (D) Measurement of cell-binding affinity by FACS using the G1 cell line.
Feng, M., Gao, W., Wang, R., Chen, W., Man, Y. G., Figg, W. D., ... & Ho, M. (2013). Therapeutically targeting glypican-3 via a conformation-specific single-domain antibody in hepatocellular carcinoma. Proceedings of the National Academy of Sciences, 110(12), E1083-E1091.
Figure 2 Binding properties of HN3 antibody.
Measurement of cell-binding affinity by FACS using the G1 cell line.
Feng, M., Gao, W., Wang, R., Chen, W., Man, Y. G., Figg, W. D., ... & Ho, M. (2013). Therapeutically targeting glypican-3 via a conformation-specific single-domain antibody in hepatocellular carcinoma. Proceedings of the National Academy of Sciences, 110(12), E1083-E1091.
Figure 3 Binding properties of HN3 antibody.
Immunoprecipitation assay using coexpressed GPC3 N- and C-terminal fragments.
Feng, M., Gao, W., Wang, R., Chen, W., Man, Y. G., Figg, W. D., ... & Ho, M. (2013). Therapeutically targeting glypican-3 via a conformation-specific single-domain antibody in hepatocellular carcinoma. Proceedings of the National Academy of Sciences, 110(12), E1083-E1091.
Figure 4 Binding properties of HN3 antibody.
HCC tissue immunohistochemistry staining with HN3.
Feng, M., Gao, W., Wang, R., Chen, W., Man, Y. G., Figg, W. D., ... & Ho, M. (2013). Therapeutically targeting glypican-3 via a conformation-specific single-domain antibody in hepatocellular carcinoma. Proceedings of the National Academy of Sciences, 110(12), E1083-E1091.
Figure 5 The effect of yap deregulation on HN3 function.
HN3 treatment of yap-knockdown cells. Proliferation was normalized with unengineered cells without antibody treatment.
Feng, M., Gao, W., Wang, R., Chen, W., Man, Y. G., Figg, W. D., ... & Ho, M. (2013). Therapeutically targeting glypican-3 via a conformation-specific single-domain antibody in hepatocellular carcinoma. Proceedings of the National Academy of Sciences, 110(12), E1083-E1091.
Figure 6 The effect of yap deregulation on HN3 function.
HN3 treatment of cells overexpressing yap-S127A.
Feng, M., Gao, W., Wang, R., Chen, W., Man, Y. G., Figg, W. D., ... & Ho, M. (2013). Therapeutically targeting glypican-3 via a conformation-specific single-domain antibody in hepatocellular carcinoma. Proceedings of the National Academy of Sciences, 110(12), E1083-E1091.
Figure 7 The effect of yap deregulation on HN3 function.
Western blot analysis of molecular changes after HN3 treatment on yap-knockdown cells or cells overexpressing yap-S127A.
Feng, M., Gao, W., Wang, R., Chen, W., Man, Y. G., Figg, W. D., ... & Ho, M. (2013). Therapeutically targeting glypican-3 via a conformation-specific single-domain antibody in hepatocellular carcinoma. Proceedings of the National Academy of Sciences, 110(12), E1083-E1091.
Figure 8 Inhibition of HCC xenograft tumor growth by HN3 antibody.
Pharmacokinetics of HN3 antibody. Serum concentrations of the antibody were measured by ELISA following a single i.v. injection of 3 mg/kg. The serum antibody concentration data were fitted with a two-compartment model using Phoenix WinNonlin software. The observed mean serum antibody concentration (in micrograms per milliliter) ± SD of four animals for each time point was plotted as a function of time.
Feng, M., Gao, W., Wang, R., Chen, W., Man, Y. G., Figg, W. D., ... & Ho, M. (2013). Therapeutically targeting glypican-3 via a conformation-specific single-domain antibody in hepatocellular carcinoma. Proceedings of the National Academy of Sciences, 110(12), E1083-E1091.
Figure 9 Inhibition of HCC xenograft tumor growth by HN3 antibody.
Inhibition of HepG2 cells inoculated into nude mice. After the tumor size reached 100 mm³, HN3 antibody or hIgG control was delivered i.v. at 60 mg/kg body weight, twice a week. Arrows indicate antibody injections.
Feng, M., Gao, W., Wang, R., Chen, W., Man, Y. G., Figg, W. D., ... & Ho, M. (2013). Therapeutically targeting glypican-3 via a conformation-specific single-domain antibody in hepatocellular carcinoma. Proceedings of the National Academy of Sciences, 110(12), E1083-E1091.
Figure 10 Inhibition of HCC xenograft tumor growth by HN3 antibody.
Inhibition of Huh-7 cells inoculated into nude mice. After the tumor size reached 100 mm³, HN3 antibody or hIgG control was delivered i.v. at 60 mg/kg body weight, twice a week. Arrows indicate antibody injections.
Feng, M., Gao, W., Wang, R., Chen, W., Man, Y. G., Figg, W. D., ... & Ho, M. (2013). Therapeutically targeting glypican-3 via a conformation-specific single-domain antibody in hepatocellular carcinoma. Proceedings of the National Academy of Sciences, 110(12), E1083-E1091.
Figure 11 Inhibition of HCC xenograft tumor growth by HN3 antibody.
Western blot analysis of molecular changes in HN3-treated and control tumors.
Feng, M., Gao, W., Wang, R., Chen, W., Man, Y. G., Figg, W. D., ... & Ho, M. (2013). Therapeutically targeting glypican-3 via a conformation-specific single-domain antibody in hepatocellular carcinoma. Proceedings of the National Academy of Sciences, 110(12), E1083-E1091.
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(Creative Biolabs Cat# TAB-416CQ, RRID: AB_3111929)
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