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Development and evaluation of detective methodology for cytotoxic activity of an anti-human epidermal growth factor receptor-2-MCC-DM1 against breast cancer cells
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Abstract:
Objective: To develop and evaluate the methodology for biological assessment of anti-HER2-MCC-DM1, an antibody-drug-conjugate (ADC) consisting of an anti-human epidermal growth factor receptor 2 (HER2) monoclonal antibody conjugated to the maytansinoid emtansine (DM1) via nonreducible thioether linkage (MCC) in breast cancer cells. Methods: Representative samples were collected from three batches of bulk drug and three batches of final products, along with the manufacturer’s validated reference standard. Breast cancer BT-474 cells were treated with the test samples and reference standard respectively. Five days after treatment, cell proliferation inhibition and cell viability were determined by colorimetric assay using a commercial cell counting kit and dual fluorescent staining analysis. Experiments were repeated three times. Results: Both test samples and reference standard of anti-HER2-MCC-DM1 inhibited BT-474 cell proliferation in a dose-dependent manner. The variation coefficient of percent inhibition within the three experiments was less than 15% for both test samples and reference standard. Compared with the nude anti-HER2 antibody, the bulk drug and final product increased BT-474 cell proliferation inhibition by （326.72±21.58）% and （315.76±34.90）% respectively. HER2-MCC-DM1 effectively induced BT-474 cell shrinkage and death as revealed by both light microscopy and fluorescence microscopy. Conclusion: The biological assessment of anti-HER2-MCC-DM1 in breast cancer BT-474 cells in vitro, developed in this study, is highly reproducible and accurate, thus offering a promising method for quality control and evaluation of ADCs.
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Project Supported:
Project supported by the National Science and Technology Major Projects for “Major New Drug Innovation and Development” (No.2014ZX09304311-001, No.2012ZX09304010)
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