Antibody Information
General Information of This Antibody
| Antibody ID | ANTI0FWHIW |
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| Antibody Name | B2C4 |
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| Organization | Anhui University of Chinese Medicine.; Biocytogen Pharmaceuticals (Beijing) Co., Ltd.; Yangtze Delta Drug Advanced Research Institute. |
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| Synonyms |
B2C4
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| Antibody Type | Bispecific antibody (BsAb) |
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| Antigen Name | Epidermal growth factor receptor (EGFR); Receptor tyrosine-protein kinase erbB-2 (ERBB2) |
Antigen Info | ||||
| Click to Show/Hide the Sequence Information of This Antibody | ||||||
The Activity Data of This Antibody
| Antibody Activity Information 1 | [1] | |||||
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Half Maximal Effective Concentration (EC50)
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0.04
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ug/mL
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NCI-H1975 cells | CVCL_1511 | ||
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| Antigen Expression | Moderate HER2 expression (HER2++); High EGFR expression (EGFR +++) | |||||
| Antibody Function | B2C4 bound to NCI-H1975 cells | |||||
| Antibody Antigen Binding Assay | For the binding assay, cells were incubated with antibodies for 30 min at 4 °C, followed by incubation with Alexa Fluor® 647 AffiniPure™ F (ab')2 Fragment Goat Anti-Human IgG antibody (Cat# 109-606-170, Jackson) for 15 min at 4 °C. For the internalization assay by fluorescence-activated cell sorting (FACS), the antibodies and pHAb-Fab (an anti-human Fab labeled with the pH-sensitive dye pHAb) were mixed in a 1:1 ratio and incubated for 15 min at room temperature in the dark to measure the internalization [27]. For Incucyte analysis, SK-OV-3 and A-431 cells were seeded at 6 × 10<sup>3</sup> or 1 × 10<sup>4</sup> cells/well and cultured overnight. After medium removal, a fresh complete medium and a mixture of antibodies and Incucyte Fabfluor-pH Antibody Labeling Dyes were added as previously described. The cells were then incubated and subjected to continuous imaging under Phase and Red conditions. | |||||
| Antibody Activity Information 2 | [1] | |||||
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Half Maximal Effective Concentration (EC50)
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0.04
|
ug/mL
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NCI-H1650 cells | CVCL_1483 | ||
| Antigen Expression | Moderate HER2 expression (HER2++); High EGFR expression (EGFR +++) | |||||
| Antibody Function | B2C4 bound to NCI-H1650 cells | |||||
| Antibody Antigen Binding Assay | For the binding assay, cells were incubated with antibodies for 30 min at 4 °C, followed by incubation with Alexa Fluor® 647 AffiniPure™ F (ab')2 Fragment Goat Anti-Human IgG antibody (Cat# 109-606-170, Jackson) for 15 min at 4 °C. For the internalization assay by fluorescence-activated cell sorting (FACS), the antibodies and pHAb-Fab (an anti-human Fab labeled with the pH-sensitive dye pHAb) were mixed in a 1:1 ratio and incubated for 15 min at room temperature in the dark to measure the internalization [27]. For Incucyte analysis, SK-OV-3 and A-431 cells were seeded at 6 × 10<sup>3</sup> or 1 × 10<sup>4</sup> cells/well and cultured overnight. After medium removal, a fresh complete medium and a mixture of antibodies and Incucyte Fabfluor-pH Antibody Labeling Dyes were added as previously described. The cells were then incubated and subjected to continuous imaging under Phase and Red conditions. | |||||
| Antibody Activity Information 3 | [1] | |||||
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Half Maximal Effective Concentration (EC50)
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0.07
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ug/mL
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SKOV-3 cells | CVCL_0532 | ||
| Antigen Expression | High HER2 expression (HER2 +++); Moderate EGFR expression (EGFR++) | |||||
| Antibody Function | B2C4 bound to SKOV-3 cells | |||||
| Antibody Antigen Binding Assay | For the binding assay, cells were incubated with antibodies for 30 min at 4 °C, followed by incubation with Alexa Fluor® 647 AffiniPure™ F (ab')2 Fragment Goat Anti-Human IgG antibody (Cat# 109-606-170, Jackson) for 15 min at 4 °C. For the internalization assay by fluorescence-activated cell sorting (FACS), the antibodies and pHAb-Fab (an anti-human Fab labeled with the pH-sensitive dye pHAb) were mixed in a 1:1 ratio and incubated for 15 min at room temperature in the dark to measure the internalization [27]. For Incucyte analysis, SK-OV-3 and A-431 cells were seeded at 6 × 10<sup>3</sup> or 1 × 10<sup>4</sup> cells/well and cultured overnight. After medium removal, a fresh complete medium and a mixture of antibodies and Incucyte Fabfluor-pH Antibody Labeling Dyes were added as previously described. The cells were then incubated and subjected to continuous imaging under Phase and Red conditions. | |||||
| Antibody Activity Information 4 | [1] | |||||
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Half Maximal Effective Concentration (EC50)
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0.24
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ug/mL
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Panc_02_03 cells | CVCL_1633 | ||
| Antigen Expression | Low HER2 expression (HER2+); Moderate EGFR expression (EGFR++) | |||||
| Antibody Function | B2C4 bound to Panc_02_03 cells | |||||
| Antibody Antigen Binding Assay | For the binding assay, cells were incubated with antibodies for 30 min at 4 °C, followed by incubation with Alexa Fluor® 647 AffiniPure™ F (ab')2 Fragment Goat Anti-Human IgG antibody (Cat# 109-606-170, Jackson) for 15 min at 4 °C. For the internalization assay by fluorescence-activated cell sorting (FACS), the antibodies and pHAb-Fab (an anti-human Fab labeled with the pH-sensitive dye pHAb) were mixed in a 1:1 ratio and incubated for 15 min at room temperature in the dark to measure the internalization [27]. For Incucyte analysis, SK-OV-3 and A-431 cells were seeded at 6 × 10<sup>3</sup> or 1 × 10<sup>4</sup> cells/well and cultured overnight. After medium removal, a fresh complete medium and a mixture of antibodies and Incucyte Fabfluor-pH Antibody Labeling Dyes were added as previously described. The cells were then incubated and subjected to continuous imaging under Phase and Red conditions. | |||||
| Antibody Activity Information 5 | [1] | |||||
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Half Maximal Effective Concentration (EC50)
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0.25
|
ug/mL
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A431 cells | CVCL_0037 | ||
| Antigen Expression | Low HER2 expression (HER2+); High EGFR expression (EGFR +++) | |||||
| Antibody Function | B2C4 bound to A431 cells | |||||
| Antibody Antigen Binding Assay | For the binding assay, cells were incubated with antibodies for 30 min at 4 °C, followed by incubation with Alexa Fluor® 647 AffiniPure™ F (ab')2 Fragment Goat Anti-Human IgG antibody (Cat# 109-606-170, Jackson) for 15 min at 4 °C. For the internalization assay by fluorescence-activated cell sorting (FACS), the antibodies and pHAb-Fab (an anti-human Fab labeled with the pH-sensitive dye pHAb) were mixed in a 1:1 ratio and incubated for 15 min at room temperature in the dark to measure the internalization [27]. For Incucyte analysis, SK-OV-3 and A-431 cells were seeded at 6 × 10<sup>3</sup> or 1 × 10<sup>4</sup> cells/well and cultured overnight. After medium removal, a fresh complete medium and a mixture of antibodies and Incucyte Fabfluor-pH Antibody Labeling Dyes were added as previously described. The cells were then incubated and subjected to continuous imaging under Phase and Red conditions. | |||||
Each Antibody-drug Conjugate Related to This Antibody
Full Information of The Activity Data of The ADC(s) Related to This Antibody
B2C4-MMAE [Investigative]
Revealed Based on the Cell Line Data
| Experiment 1 Reporting the Activity Date of This ADC | [1] | ||||
| Efficacy Data | Half Maximal inhibitory Concentration (lC50) |
0.02 ug/mL
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High HER2 expression (HER2 +++); Moderate EGFR expression (EGFR++) | ||
| Method Description |
A-431 was inoculated at 6 × 103 cells/well, NCI-H1650 at 5 × 103 cells/well, SK-OV-3, Panc 02.03, and NCI-H520 at 4 × 103 cells/well, and NCI-H1975 at 3 × 103 cells/well in fresh complete medium and cultured overnight. Antibodies were diluted with a complete medium and added to the cells. The 96-well plates were placed in IncuCyte for incubation and were photographed at 4-h intervals for 4 days under phase conditions. This allowed for the analysis of cell viability by measuring the confluence of the cells in each well.
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| In Vitro Model | Ovarian serous adenocarcinoma | SKOV-3 cells | CVCL_0532 | ||
| Experiment 2 Reporting the Activity Date of This ADC | [1] | ||||
| Efficacy Data | Half Maximal inhibitory Concentration (lC50) |
0.04 ug/mL
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Moderate HER2 expression (HER2++); High EGFR expression (EGFR +++) | ||
| Method Description |
A-431 was inoculated at 6 × 103 cells/well, NCI-H1650 at 5 × 103 cells/well, SK-OV-3, Panc 02.03, and NCI-H520 at 4 × 103 cells/well, and NCI-H1975 at 3 × 103 cells/well in fresh complete medium and cultured overnight. Antibodies were diluted with a complete medium and added to the cells. The 96-well plates were placed in IncuCyte for incubation and were photographed at 4-h intervals for 4 days under phase conditions. This allowed for the analysis of cell viability by measuring the confluence of the cells in each well.
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| In Vitro Model | Lung adenocarcinoma | NCI-H1650 cells | CVCL_1483 | ||
| Experiment 3 Reporting the Activity Date of This ADC | [1] | ||||
| Efficacy Data | Half Maximal inhibitory Concentration (lC50) |
0.05 ug/mL
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Low HER2 expression (HER2+); High EGFR expression (EGFR +++) | ||
| Method Description |
A-431 was inoculated at 6 × 103 cells/well, NCI-H1650 at 5 × 103 cells/well, SK-OV-3, Panc 02.03, and NCI-H520 at 4 × 103 cells/well, and NCI-H1975 at 3 × 103 cells/well in fresh complete medium and cultured overnight. Antibodies were diluted with a complete medium and added to the cells. The 96-well plates were placed in IncuCyte for incubation and were photographed at 4-h intervals for 4 days under phase conditions. This allowed for the analysis of cell viability by measuring the confluence of the cells in each well.
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| In Vitro Model | Skin squamous cell carcinoma | A431 cells | CVCL_0037 | ||
| Experiment 4 Reporting the Activity Date of This ADC | [1] | ||||
| Efficacy Data | Half Maximal inhibitory Concentration (lC50) |
0.05 ug/mL
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Moderate HER2 expression (HER2++); High EGFR expression (EGFR +++) | ||
| Method Description |
A-431 was inoculated at 6 × 103 cells/well, NCI-H1650 at 5 × 103 cells/well, SK-OV-3, Panc 02.03, and NCI-H520 at 4 × 103 cells/well, and NCI-H1975 at 3 × 103 cells/well in fresh complete medium and cultured overnight. Antibodies were diluted with a complete medium and added to the cells. The 96-well plates were placed in IncuCyte for incubation and were photographed at 4-h intervals for 4 days under phase conditions. This allowed for the analysis of cell viability by measuring the confluence of the cells in each well.
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| In Vitro Model | Lung adenocarcinoma | NCI-H1975 cells | CVCL_1511 | ||
| Experiment 5 Reporting the Activity Date of This ADC | [1] | ||||
| Efficacy Data | Half Maximal inhibitory Concentration (lC50) |
0.29 ug/mL
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Low HER2 expression (HER2+); Moderate EGFR expression (EGFR++) | ||
| Method Description |
A-431 was inoculated at 6 × 103 cells/well, NCI-H1650 at 5 × 103 cells/well, SK-OV-3, Panc 02.03, and NCI-H520 at 4 × 103 cells/well, and NCI-H1975 at 3 × 103 cells/well in fresh complete medium and cultured overnight. Antibodies were diluted with a complete medium and added to the cells. The 96-well plates were placed in IncuCyte for incubation and were photographed at 4-h intervals for 4 days under phase conditions. This allowed for the analysis of cell viability by measuring the confluence of the cells in each well.
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| In Vitro Model | Pancreatic adenocarcinoma | Panc_02_03 cells | CVCL_1633 | ||
| Experiment 6 Reporting the Activity Date of This ADC | [1] | ||||
| Efficacy Data | Half Maximal inhibitory Concentration (lC50) |
4.16 ug/mL
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Negative HER2 expression (HER2-); Negative EGFR expression (EGFR-) | ||
| Method Description |
A-431 was inoculated at 6 × 103 cells/well, NCI-H1650 at 5 × 103 cells/well, SK-OV-3, Panc 02.03, and NCI-H520 at 4 × 103 cells/well, and NCI-H1975 at 3 × 103 cells/well in fresh complete medium and cultured overnight. Antibodies were diluted with a complete medium and added to the cells. The 96-well plates were placed in IncuCyte for incubation and were photographed at 4-h intervals for 4 days under phase conditions. This allowed for the analysis of cell viability by measuring the confluence of the cells in each well.
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| In Vitro Model | Lung squamous cell carcinoma | NCI-H520 cells | CVCL_1566 | ||
References