The Antibody-Drug Conjugate (ADC) structure comprises low-molecular-weight drugs bound to the cysteine or lysine residues of monoclonal IgG. Currently, ADCs are attracting attention as a next-generation drug that combines the advantages of small molecule weight drugs and antibody drugs. Drug Antibody Ratio (DAR) is an important evaluation point for quality control of the ADC. A polymer-based reverse phase chromatography column ODP2 HP-4B was used in hydrophobic chromatography mode. In conclusion, commercially available cysteine-linked ADCs can be separated in ascending order of DAR with ODP2 HP-4B.
*The number after DAR in the figure indicates the number of low-molecular-weight drug bonds.
Sample: 5 µL
Sigma MAb Antibody-Drug Conjugate (ADC)
ADC Mimic 1 mg/mL (in H2O)
Column : Shodex ODP2 HP-4B (4.6 mm I.D. x 50 mm)
Eluent : (A) 1.5 M (NH4)2SO4 in 50 mM Sodium phosphate buffer (pH7.0)
(B) 50 mM Sodium phosphate buffer (pH7.0)/IPA=75/25
High pressure linear gradient;
(B %) 0 % to 100 % (0 to 20 min), 100 % (20 to 30 min),
100 to 0 % (30 to 30.01 min), 0 % (30.01 to 35 min)
Flow rate : 0.8 mL/min
Detector : UV (280 nm)
Column temp. : 30 °C
