UHPLC Analysis of Glembatumumab Vedotin Antibody Drug Conjugate on BIOshell(TM) A400 Protein C4

UHPLC Analysis of Glembatumumab Vedotin Antibody Drug Conjugate on BIOshell™ A400 Protein C4


column BIOshell A400 Protein C4; 10 cm x 2.1 mm I.D., 3.4 μm particles (66825-U)
column temp. 90 °C
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)
sample Glembatumumab vedotin, 355 μg/mL, 0.1% TFA in water
injection 1.0 μL
detector UV, 215 nm


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.
Featured Industry Life Science and Biopharma
Legal Information BIOshell is a trademark of Sigma-Aldrich Co. LLC
suitability application for UHPLC


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