Comparison of retention time of DAR species in RP-HPLC

DAR speciesTrastuzumab-MCC-maytansinoidTrastuzumab-MC-VC-PAB-MMAE
LC7.0 min7.0 min
LC + drug-linker8.2 min9.6 min
HC10.2 min10.2 min
HC + drug-linker11.0 min12.1 min
HC + 2 drug-linkers13.1 min15.4 min
HC + 3 drug-linkers14.4 min18.0 min

LC: antibody light chain; LC + drug-linker: LC conjugated with one drug-linker; HC + drug-linker: HC conjugated with one drug-linker