Drug Antibody Ratio Analysis of Antibody-Drug Conjugate (ODP2 HP-4B)

Antibody-Drug Conjugate (ADC) has a structure in which low-molecular-weight drugs are bound to the cysteine or lysine residues of monoclonal IgG. ADCs are attracting attention as a next-generation drug that combines the advantages of small molecule weight drugs and antibody drugs. In ADC, Drug Antibody Ratio (DAR) is one of the important evaluation points for quality control. Using a polymer-based reverse phase chromatography column ODP2 HP-4B  in hydrophobic chromatography mode, it was confirmed that commercially available cysteine-linked ADCs can be separated in ascending order of DAR.
*The number after DAR in the figure indicates the number of low-molecular-weight drug bonds.


Sample : 5 µL
SigmaMAb 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

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