LC及びMS技術を用いたADCの薬物抗体比及び薬物結合位置の解析 / The Characterization of Drug-to-Antibody Ratio and Isoforms of ADCs Using LC and MS Technologies
- Resource Type
- Journal Article
- Authors
- April Xu; Frank Kotch; Henry Shion; Reiko Iizuka; Robert Birdsall; Thomas Porter; Weibin Chen; 飯塚 玲子
- Source
- YAKUGAKU ZASSHI. 2017, 137(5):525
- Subject
- LC/MS
antibody-drug conjugate
critical quality attributes
drug-to-antibody ratio
hydrophobic interaction chromatography
- Language
- Japanese
- ISSN
- 0031-6903
1347-5231
Hydrophobic interaction chromatography (HIC) and MS are leading techniques for the characterization of the critical quality attributes (CQA) of antibody-drug conjugates (ADCs). This includes the average drug-to-antibody ratio (DAR) and drug loading distribution. A workflow that effectively utilizes the synergy between chromatography and detection technologies has been developed and was assessed using cysteine-conjugated ADCs. The DAR of low, moderate and high drug-loaded ADC samples were calculated from the chromatographic peak areas using LC(HIC)/UV or the deconvoluted mass spectra using native LC(SEC)/MS. The results of DAR by both technologies produced comparable results. In addition, the 2D-LC/MS system has been evaluated in combination with HIC and reversed-phase chromatography for structural identification. Individual peaks from the 1st dimension of the HIC separation were isolated online and re-directed to the 2nd dimension reversed-phase column. ADC was detected as the sub-units by MS and the conjugation site was identified via a middle down approach.