UHPLC Analysis of Glembatumumab Vedotin Antibody Drug Conjugate on BIOshell A400 Protein C4
CONDITIONS
column
BIOshell A400 Protein C4; 10 cm x 2.1 mm I.D., 3.4 μm particles (66825-U)
mobile phase
[A] 95:5 (0.1% TFA in water: 0.1% TFA in n-propanol); [B] 70:20:10 (0.1% TFA in n-propanol: 0.1% TFA in acetonitrile: 0.1% TFA in water)
gradient
15% B to 25% B in 20 min
flow rate
0.2 mL/min
pressure
650 psi (44.8 bar)
column temp.
90 °C
detector
UV, 215 nm
injection
1.0 μL
sample
Glembatumumab vedotin, 355 μg/mL, 0.1% TFA in water
Description
Analysis Note
Glembatumumab vedotin is an antibody-drug conjugate (ADC) that targets cancer cells expressing transmembrane glycoprotein NMB. This ADC is one approved therapeutic to treat triple negative breast cancer. Glembatumumab is an IgG2 kappa monoclonal antibody; therefore, there are six different interchain disulfide bonds, which can lead to 12 different ADC species. This is further complicated by there being three additional isomers of IgG2 that can also be conjugated with cytotoxic payload. Such a heterogeneous sample can make quantitation of the correct dose form of the ADC difficult. This application demonstrates the ability of the BIOshell A400 Protein C4 column to provide resolution of impurities and several different conjugated species.
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