문수경 (안전성평가연구소)

문수경 (안전성평가연구소)

문수경 (안전성평가연구소)

김형민 (충남대학교)

강종성 (충남대학교)

민정은 (안전성평가연구소)

엄한영 (안전성평가연구소)

조현덕 (안전성평가연구)

Antibody-drug conjugates (ADCs) are innovative therapeutics that combine the benefits of antibodies and cytotoxic drugs by linking the cytotoxic agent to an antibody. This study developed a unified quantification method for the pharmacokinetic evaluation of ADCs using LC-MS analysis, focusing on three parameters: total antibody, antibody-drug conjugate (acDrug), and free payload. The analyte used for this study was trastuzumab deruxtecan (T-Dxd, Enhertu^®), a treatment for HER2-positive breast cancer. For total antibody analysis, an immunocapture technique was employed to extract the target component, followed by trypsin digestion to analyze the signature peptide. To analyze acDrug and free payload, a method was established to detect Dxd, the cytotoxic drug of T-Dxd in rat plasma. The acDrug analysis involved immunocapture followed by enzymatic cleavage of the linker for quantification. These methods were validated according to FDA and ICH bioanalytical method validation guidelines. It met the criteria for system suitability, linearity, accuracy, precision, carry-over, and recovery, demonstrating its validity and reliability. By adhering to regulatory guidelines, this unified LC-MS-based quantification method offers enhanced analytical efficiency for the bioanalysis, pharmacokinetic/pharmacodynamic (PK/PD) studies, and various pharmaceutical research applications of ADC.