하정엽 (오송첨단의료산업진흥재단)

진종화 (오송첨단의료산업진흥재단)

하정엽 (오송첨단의료산업진흥재단)

이경욱 (오송첨단의료산업진흥재단)

이민영 (오송첨단의료산업진흥재단)

노영욱 (오송첨단의료산업진흥재단)

진종화 (오송첨단의료산업진흥재단)

Trastuzumab deruxtecan (Enhertu®, T-Dxd) is a HER2-targeting antibody-drug conjugate (ADC) with a high drug-to-antibody ratio (DAR ~8) and a tumor-selective release mechanism. Due to its structural complexity, rigorous analytical workflows are required to evaluate product quality and linker stability throughout development.

In this study, we developed an integrated analytical platform combining SEC-HPLC, HIC-HPLC, and LC-MS/MS. The physicochemical properties including monomer content, aggregation, and DAR distribution were assessed using HPLC methods, while batch homogeneity and conjugation profiles were confirmed via intact mass analysis. A reference batch was selected for in vitro and in vivo stability testing.

Linker stability was assessed by monitoring DXd release from T-Dxd incubated in mouse serum over time and by quantifying free DXd in mouse plasma following intravenous dosing. Results indicated time-dependent cleavage of the linker, suggesting partial instability under physiological conditions.

This platform enables comprehensive ADC characterization and supports its application in pharmacokinetic interpretation, formulation development, and lot-release testing.