General Information of This Antibody
Antibody ID
ANTI0FQTUS
Antibody Name
C4
Organization
Anhui University of Chinese Medicine.; Biocytogen Pharmaceuticals (Beijing) Co., Ltd.; Yangtze Delta Drug Advanced Research Institute.
Synonyms
C4
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Antibody Type
Monoclonal antibody (mAb)
Antibody Subtype
Murine lgG1
Antigen Name
Epidermal growth factor receptor (EGFR)
 Antigen Info 
Click to Show/Hide the Sequence Information of This Antibody
The Activity Data of This Antibody
Antibody Activity Information 1 [1]
Half Maximal Effective Concentration (EC50)
0.03
ug/mL
SKOV-3 cells CVCL_0532 
Antigen Expression Moderate EGFR expression (EGFR++)
Antibody Function C4 bound to SKOV-3 cells
Antibody Antigen Binding Assay For the binding assay, cells were incubated with antibodies for 30 min at 4 &deg;C, followed by incubation with Alexa Fluor&reg; 647 AffiniPure&trade; F (ab')2 Fragment Goat Anti-Human IgG antibody (Cat# 109-606-170, Jackson) for 15 min at 4 &deg;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 &times; 10<sup>3</sup> or 1 &times; 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]
Half Maximal Effective Concentration (EC50)
0.03
ug/mL
NCI-H1975 cells CVCL_1511 
Antigen Expression High EGFR expression (EGFR +++)
Antibody Function C4 bound to NCI-H1975 cells
Antibody Antigen Binding Assay For the binding assay, cells were incubated with antibodies for 30 min at 4 &deg;C, followed by incubation with Alexa Fluor&reg; 647 AffiniPure&trade; F (ab')2 Fragment Goat Anti-Human IgG antibody (Cat# 109-606-170, Jackson) for 15 min at 4 &deg;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 &times; 10<sup>3</sup> or 1 &times; 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]
Half Maximal Effective Concentration (EC50)
0.05
ug/mL
NCI-H1650 cells CVCL_1483 
Antigen Expression High EGFR expression (EGFR +++)
Antibody Function C4 bound to NCI-H1650 cells
Antibody Antigen Binding Assay For the binding assay, cells were incubated with antibodies for 30 min at 4 &deg;C, followed by incubation with Alexa Fluor&reg; 647 AffiniPure&trade; F (ab')2 Fragment Goat Anti-Human IgG antibody (Cat# 109-606-170, Jackson) for 15 min at 4 &deg;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 &times; 10<sup>3</sup> or 1 &times; 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]
Half Maximal Effective Concentration (EC50)
0.1
ug/mL
Panc_02_03 cells CVCL_1633 
Antigen Expression Moderate EGFR expression (EGFR++)
Antibody Function C4 bound to Panc_02_03 cells
Antibody Antigen Binding Assay For the binding assay, cells were incubated with antibodies for 30 min at 4 &deg;C, followed by incubation with Alexa Fluor&reg; 647 AffiniPure&trade; F (ab')2 Fragment Goat Anti-Human IgG antibody (Cat# 109-606-170, Jackson) for 15 min at 4 &deg;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 &times; 10<sup>3</sup> or 1 &times; 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]
Half Maximal Effective Concentration (EC50)
0.33
ug/mL
A431 cells CVCL_0037 
Antigen Expression High EGFR expression (EGFR +++)
Antibody Function C4 bound to A431 cells
Antibody Antigen Binding Assay For the binding assay, cells were incubated with antibodies for 30 min at 4 &deg;C, followed by incubation with Alexa Fluor&reg; 647 AffiniPure&trade; F (ab')2 Fragment Goat Anti-Human IgG antibody (Cat# 109-606-170, Jackson) for 15 min at 4 &deg;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 &times; 10<sup>3</sup> or 1 &times; 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
C4-MMAE [Investigative]
Revealed Based on the Cell Line Data
Click To Hide/Show 11 Activity Data Related to This Level
Experiment 1 Reporting the Activity Date of This ADC [2]
Efficacy Data Half Maximal inhibitory Concentration (lC50)
0.54 uM
Moderate HER2 expression (HER2++)
Method Description
The in vitro antitumor efficacy of ADC was assessed using the BxPc-3 cell.
In Vitro Model Pancreatic ductal adenocarcinoma BxPC-3 cells CVCL_0186
Experiment 2 Reporting the Activity Date of This ADC [2]
Efficacy Data Half Maximal inhibitory Concentration (lC50)
0.54 uM
Low HER2 expression (HER2+)
Method Description
The in vitro antitumor efficacy of ADC was assessed using the MDA-MB-468 cell.
In Vitro Model Breast adenocarcinoma MDA-MB-468 cells CVCL_0419
Experiment 3 Reporting the Activity Date of This ADC [2]
Efficacy Data Half Maximal inhibitory Concentration (lC50)
0.85 uM
Low HER2 expression (HER2+)
Method Description
The in vitro antitumor efficacy of ADC was assessed using the MDA-MB-231 cell.
In Vitro Model Breast adenocarcinoma MDA-MB-231 cells CVCL_0062
Experiment 4 Reporting the Activity Date of This ADC [2]
Efficacy Data Half Maximal inhibitory Concentration (lC50)
3.8 uM
Low HER2 expression (HER2+)
Method Description
The in vitro antitumor efficacy of ADC was assessed using the AsPc1 cell.
In Vitro Model Pancreatic ductal adenocarcinoma AsPc1 cells CVCL_0152
Experiment 5 Reporting the Activity Date of This ADC [2]
Efficacy Data Half Maximal inhibitory Concentration (lC50)
10.79 uM
Moderate HER2 expression (HER2++)
Method Description
The in vitro antitumor efficacy of ADC was assessed using the CFPAC cell.
In Vitro Model Cystic fibrosis, Pancreatic ductal adenocarcinoma CFPAC cells CVCL_1119
Experiment 6 Reporting the Activity Date of This ADC [1]
Efficacy Data Half Maximal inhibitory Concentration (lC50)
0.01 ug/mL
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 7 Reporting the Activity Date of This ADC [1]
Efficacy Data Half Maximal inhibitory Concentration (lC50)
0.02 ug/mL
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 8 Reporting the Activity Date of This ADC [1]
Efficacy Data Half Maximal inhibitory Concentration (lC50)
0.02 ug/mL
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 9 Reporting the Activity Date of This ADC [1]
Efficacy Data Half Maximal inhibitory Concentration (lC50)
0.05 ug/mL
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 10 Reporting the Activity Date of This ADC [1]
Efficacy Data Half Maximal inhibitory Concentration (lC50)
0.12 ug/mL
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 11 Reporting the Activity Date of This ADC [1]
Efficacy Data Half Maximal inhibitory Concentration (lC50)
4.18 ug/mL
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
Ref 1 A novel anti-HER2/EGFR bispecific antibody-drug conjugate demonstrates promising antitumor efficacy and overcomes resistance to HER2- or EGFR-targeted ADCs
Ref 2 A comparison of the activity, lysosomal stability, and efficacy of legumain-cleavable and cathepsin-cleavable ADC linkers