Anti-Human HER2/neu Recombinant Antibody (HuA21)

The inhibitory effects of HuA21, trastuzumab (Tra), or the combination on the growth of the: BT-474 (A); BT-474/HR (B); and SKBR3 (C) cells. The cells were incubated with HuA21 or Tra at the indicated concentrations for 72 h. Cell viability was measured by the the cell counting kit-8 (CCK-8) assay. The curve charts represent the cell viability of the: BT-474 (A); BT-474/HR (B); and SKBR3 cells (C). The experiments are performed in triplicate, and the data are representative of three separate experiments. # p < 0.05 vs. control; & p < 0.05 vs. HuA21-treated cells.

Li, R., Hu, S., Chang, Y., Zhang, Z., Zha, Z., Huang, H., ... & Wei, W. (2016). Development and characterization of a humanized anti-HER2 antibody HuA21 with potent anti-tumor properties in breast cancer cells. International journal of molecular sciences, 17(4), 563.

HuA21-mediated ADCC against HER2-positive breast cancer cells BT-474 cells were pre-incubated with the indicated concentrations of HuA21 or trastuzumab (Tra). The ADCC experiments were performed in triplicate, and the data are representative of three separate experiments.

Li, R., Hu, S., Chang, Y., Zhang, Z., Zha, Z., Huang, H., ... & Wei, W. (2016). Development and characterization of a humanized anti-HER2 antibody HuA21 with potent anti-tumor properties in breast cancer cells. International journal of molecular sciences, 17(4), 563.

The representative images and bar graph indicate the inhibitory effects of HuA21 (10 µg/mL), trastuzumab (Tra, 10 µg/mL), or HuA21 (10 µg/mL) + Tra (10 µg/mL) on the migration of SKBR3 breast cancer cells. The cells were treated with HuA21, Tra, or HuA21 + Tra for 24 h. Magnification: 100×. * p < 0.05 vs. control; & p < 0.05 vs. HuA21-treated cells. The experiments were performed in triplicate, and the data are representative of three separate experiments.

Li, R., Hu, S., Chang, Y., Zhang, Z., Zha, Z., Huang, H., ... & Wei, W. (2016). Development and characterization of a humanized anti-HER2 antibody HuA21 with potent anti-tumor properties in breast cancer cells. International journal of molecular sciences, 17(4), 563.

Western blot and the representative picture showing the bands of EGFR, p-HER2, total HER2, total HER3, p-Akt, total Akt, p-Erk1/2, and total Erk1/2 expressed in the BT-474 treated with HuA21, trastuzumab (Tra), or HuA21 + Tra. β-Actin was used as the loading control. * p < 0.05 vs. control; & p < 0.05 vs. HuA21-treated cells. The experiments were performed in triplicate, and the data are representative of three separate experiments.

Li, R., Hu, S., Chang, Y., Zhang, Z., Zha, Z., Huang, H., ... & Wei, W. (2016). Development and characterization of a humanized anti-HER2 antibody HuA21 with potent anti-tumor properties in breast cancer cells. International journal of molecular sciences, 17(4), 563.

The BT-474cells were incubated with FITC-conjugated HuA21 or FITC-conjugated trastuzumab (Tra) for the indicated time periods, and then the presence of HuA21 or Tra was revealed by fluorescence microscope. Representative photomicrographs showing HuA21 or Tra (Green: FITC-conjugated HuA21 or Tra, scale bar = 50 µm).

Li, R., Hu, S., Chang, Y., Zhang, Z., Zha, Z., Huang, H., ... & Wei, W. (2016). Development and characterization of a humanized anti-HER2 antibody HuA21 with potent anti-tumor properties in breast cancer cells. International journal of molecular sciences, 17(4), 563.

The BT-474 cells were incubated with HuA21 or Tra for 4 h, FITC-conjugated secondary antibody was added and the signal was detected by confocal microscopy (Green: FITC-conjugated HuA21 or Tra; nuclei (blue) was stained by 4',6-diamidino-2-phenylindole (DAPI), scale bar = 50 µm)

Li, R., Hu, S., Chang, Y., Zhang, Z., Zha, Z., Huang, H., ... & Wei, W. (2016). Development and characterization of a humanized anti-HER2 antibody HuA21 with potent anti-tumor properties in breast cancer cells. International journal of molecular sciences, 17(4), 563.

The BT-474cells were incubated with Tra or HuA21 for the indicated periods. HuA21 or Tra on the cellsurface was labeled with a FITC-conjugated secondary antibody and analyzed by flow cytometry.

Li, R., Hu, S., Chang, Y., Zhang, Z., Zha, Z., Huang, H., ... & Wei, W. (2016). Development and characterization of a humanized anti-HER2 antibody HuA21 with potent anti-tumor properties in breast cancer cells. International journal of molecular sciences, 17(4), 563.