Development and Validation of the ELISA Method for Neutralizing Anti-trastuzumab Antibodies Detection in Human Blood Serum

Introduction. Trastuzumab is the first known anti-HER2 agent, which revolutionized treatment of one most common cancer types – breast cancer. Despite trastuzumab being approved long time ago, further improvement related analytical methods remains relevant primarily due to emergence new biosimilars. For instance, immunogenicity adverse reaction usually associated with biological drugs, can still be for trastuzumab. Anti-drug antibodies, including neutralizing caused by therapy, affect drug effectiveness and safety profile. Aim. The aim this study was develop validate method anti-trastuzumab antibodies determination in human blood serum. Materials methods. antibody carried out competitive ELISA method, using spectrophotometric detection visible range spectrum. Results discussion. developed validated cut-point, selectivity, sensitivity, specificity, precision stability (short-term long-term). To decrease background noise from non-specific binding sera components, minimum required dilution value determined at 0.5 % calculated cut-point 14.62 %. sensitivity estimated 1985.2 ng/mL antibodies. Conclusion. obtained results allowed us apply serum during assessment bioequivalence clinical trials.