Human Anti-TM4SF5 Recombinant Antibody (clone hEC2-C-1) (CAT#: FAMAB-0707WJ)

Figure 1 Binding affinity of the humanized antibody for the cyclic peptide hTM4SF5EC2-C was analyzed using ELISA.

Wu, G., Kim, D., Park, B. K., Park, S., Ha, J. H., Kim, T. H., ... & Lee, Y. (2016). Anti-metastatic effect of the TM4SF5-specific peptide vaccine and humanized monoclonal antibody on colon cancer in a mouse lung metastasis model. Oncotarget, 7(48), 79170.

Figure 2 Binding affinity for the cyclic peptide was analyzed using SPR system.

Wu, G., Kim, D., Park, B. K., Park, S., Ha, J. H., Kim, T. H., ... & Lee, Y. (2016). Anti-metastatic effect of the TM4SF5-specific peptide vaccine and humanized monoclonal antibody on colon cancer in a mouse lung metastasis model. Oncotarget, 7(48), 79170.

Figure 3 Whole cell protein lysates from HEK 293F-EV and HEK 293F-TM4SF5 cells were analyzed by western blotting with humanized anti-TM4SF5 antibody (hEC2-C-2) or anti-β-actin antibody.

Whole cell protein lysates from HEK 293F-EV and HEK 293F-TM4SF5 cells were immunoprecipitated with hEC2-C-2 antibody and analyzed by western blotting with the anti-Myc antibody.

Wu, G., Kim, D., Park, B. K., Park, S., Ha, J. H., Kim, T. H., ... & Lee, Y. (2016). Anti-metastatic effect of the TM4SF5-specific peptide vaccine and humanized monoclonal antibody on colon cancer in a mouse lung metastasis model. Oncotarget, 7(48), 79170.

Figure 4 CT-26 and HCT-116 cells were analyzed by confocal microscopy using normal human IgG and hEC2-C-2 antibody.

Scale bars, 20 μm.

Wu, G., Kim, D., Park, B. K., Park, S., Ha, J. H., Kim, T. H., ... & Lee, Y. (2016). Anti-metastatic effect of the TM4SF5-specific peptide vaccine and humanized monoclonal antibody on colon cancer in a mouse lung metastasis model. Oncotarget, 7(48), 79170.

Figure 5 Effects of the humanized anti-TM4SF5 monoclonal antibody on the migration of colon cancer cells

(A) Migration assay. Scale bars, 100 μm.

Wu, G., Kim, D., Park, B. K., Park, S., Ha, J. H., Kim, T. H., ... & Lee, Y. (2016). Anti-metastatic effect of the TM4SF5-specific peptide vaccine and humanized monoclonal antibody on colon cancer in a mouse lung metastasis model. Oncotarget, 7(48), 79170.

Figure 6 Effects of the humanized anti-TM4SF5 monoclonal antibody on the migration of colon cancer cells

Wound-healing assay. Scale bars, 300 μm. Migration properties of CT-26 and HCT-116 cells were compared after treatment with PBS, normal human IgG, or hEC2-C-2 antibody.

Wu, G., Kim, D., Park, B. K., Park, S., Ha, J. H., Kim, T. H., ... & Lee, Y. (2016). Anti-metastatic effect of the TM4SF5-specific peptide vaccine and humanized monoclonal antibody on colon cancer in a mouse lung metastasis model. Oncotarget, 7(48), 79170.

Figure 7 Effect of the humanized anti-TM4SF5 antibody on the expression of adhesion molecules in colon cancer cells

CT-26 and HCT-116 cells were treated with PBS, normal human IgG, or hEC2-C-2 antibody. Expression of E-cadherin and β-catenin was determined by western blotting with anti-E-cadherin and anti-β-catenin antibodies. (A) Confocal microscopy images after 3 h of treatment with DyLight 488-labeled human IgG control or humanized anti-TM4SF5 antibody (hEC2-C-2 Ab; 10 μg/mL). Scale bars, 20 μm.

Wu, G., Kim, D., Park, B. K., Park, S., Ha, J. H., Kim, T. H., ... & Lee, Y. (2016). Anti-metastatic effect of the TM4SF5-specific peptide vaccine and humanized monoclonal antibody on colon cancer in a mouse lung metastasis model. Oncotarget, 7(48), 79170.

Figure 8 Effect of the humanized anti-TM4SF5 antibody on the expression of adhesion molecules in colon cancer cells

CT-26 and HCT-116 cells were treated with PBS, normal human IgG, or hEC2-C-2 antibody. Expression of E-cadherin and β-catenin was determined by western blotting with anti-E-cadherin and anti-β-catenin antibodies. (B) Western blot analysis. β-actin was used as an internal control. The band intensities were measured and quantitative changes are shown on the graph.

Wu, G., Kim, D., Park, B. K., Park, S., Ha, J. H., Kim, T. H., ... & Lee, Y. (2016). Anti-metastatic effect of the TM4SF5-specific peptide vaccine and humanized monoclonal antibody on colon cancer in a mouse lung metastasis model. Oncotarget, 7(48), 79170.

Figure 9 Survival rate in the lung metastasis mouse model

CT-26 cells were intravenously injected into BALB/c mice. On day 5, PBS, human IgG, or hEC2-C-2 antibody was intravenously injected into the mice, and tumor growth was monitored after 22 days (PBS controls, n = 8; colon cancer cell-injected group, n = 12 each). (A) Experimental schedule. (B) Body weights were measured every other day for 20 days. (C) Survival of the humanized anti-TM4SF5 antibody-injected mice after implantation with CT-26 cells.

Wu, G., Kim, D., Park, B. K., Park, S., Ha, J. H., Kim, T. H., ... & Lee, Y. (2016). Anti-metastatic effect of the TM4SF5-specific peptide vaccine and humanized monoclonal antibody on colon cancer in a mouse lung metastasis model. Oncotarget, 7(48), 79170.

Figure 10 Inhibition of lung metastasis by the humanized anti-TM4SF5 monoclonal antibody

CT-26 cells were intravenously injected into BALB/c mice. On day 5, PBS, human IgG, or hEC2-C-2 antibody was intravenously injected into the mice, and tumor growth was monitored after 19 days (PBS controls, n = 8; colon cancer cell-injected group, n = 12 each). (A) Experimental schedule. (B) Macroscopic appearance of the lung. (C) Weight of the mouse lungs. **P < 0.01. (D) Histological examination of the lung tissues. Scale bars: 100X, 100 μm; 400X, 25 μm.

Wu, G., Kim, D., Park, B. K., Park, S., Ha, J. H., Kim, T. H., ... & Lee, Y. (2016). Anti-metastatic effect of the TM4SF5-specific peptide vaccine and humanized monoclonal antibody on colon cancer in a mouse lung metastasis model. Oncotarget, 7(48), 79170.