Chemistry Colloquium: Using Design of Experiments for Efficient ADC Methods Development

Join the Department of Chemistry for this colloquium featuring guest speaker Gregory Webster, a senior principal research scientist in analytical research and development at Abbvie.

Liquid chromatography methods used to characterize Antibody-Drug Conjugate (ADC) formulations typically involve size exclusion chromatography (SEC), hydrophobic interaction chromatography (HIC) and both intact and denatured reversed phased chromatography (RP-LC) procedures. Each of these techniques present unique challenges when being looked at through the perspective of traditional small molecule chromatography requirements. The nature and size of the molecule hinder the protein from following “small molecule” rules while typically exhibiting a much less resolved chromatographic profile.

SEC methods are routinely used in the pharmaceutical industry to characterize the higher and lower molecular weight species of the protein. In our laboratory, column lifetime issues have been
experienced with varying column manufacturers and method conditions. This challenges not only method transferability but the usefulness of the analytical method as the molecule proceeds to regulatory approval and routine testing requirements. Typically, robustness studies for SEC methods for proteins can involve characterizing the effect of column temperature, flow rate, and whether a salt or organic modifier (typically IPA) needs to be added to the mobile phase and at what levels. These robustness parameters provide an excellent platform to evaluate protein SEC method conditions and several of the commercially available SEC columns on the market.

The challenges, opportunities and optimizations needed to execute these DOE studies and their projections of SEC method conditions for ADC formulations will be presented.