Analytical Separation of Antibody Drug Conjugates (ADCs) and Monoclonal Antibodies (mAbs) Using Various Chromatographic Methods

This webcast was recorded on September 30, 2014.


Antibody Drug Conjugates (ADCs) are more complex and heterogeneous than their corresponding unconjugated base antibody. Different chromatographic modes can be selected for analytical scale purification, based on the specificity of the individual ADC, the nature of the linker, the attachment sites, and more. With the advancement of HPLC analytical column chromatography, the separation of complex peptide mixtures derived from protease digestions of monoclonal antibodies (mAbs) and ADCs are possible. Here we present a brief overview of the separation of mAbs and ADCs using various chromatographic modes at analytical scale. In particular, we will discuss:

• HIC (Hydrophobic Interaction Chromatography), which can be used for the chromatographic analysis of drug distribution for the conjugates formed by interchain disulfide bonds.
• Charge-based separations such as ion exchange chromatography (IEC), isoelectric focusing gel electrophoresis (IEF) and capillary isoelectric focusing (cIEF), which are used to estimate the drug distribution for ADCs.
• Size exclusion chromatography (SEC), which is useful to measure the extent of aggregation and fragmentation for stability testing.


Justin Steve
Technical Service Specialist
Tosoh Bioscience LLC

Meg L'Heureux

Managing Editor

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