Technical Note : Integrated Anti-HER2 ADC Profiling: Internalization, Site-Specific Conjugation, and Cytotoxicity
Case study
Integrated Anti-HER2 ADC Profiling: Internalization, Site-Specific Conjugation, and Cytotoxicity
• Transient expression and Protein A purification of anti-HER2 candidates selected by single-cell Beacon® screening
• Internalization kinetics of naked antibodies or MMAF conjugates measured by pHSense assay (Revvity) on HER2⁺ SK-BR-3 cells
• Site-specific two-step BTG conjugation to DBCO-PEG4-MMAF; DAR = 4.0 confirmed by LC-MS
• Cytotoxicity assessed on SK-BR-3 cells by the CellTiter-Glo® Luminescent Cell Viability Assay
• Composite score (Beacon® affinity + naked antibody internalization) predicts ADC cytotoxicity with AUC = 0.852 and 100% sensitivity
Download the technical note
Integrated Anti-HER2 ADC Profiling: Internalization, Site-Specific Conjugation, and Cytotoxicity
Technical Note : Integrated Anti-HER2 ADC Profiling: Internalization, Site-Specific Conjugation, and Cytotoxicity
