Using design of experiments to optimise SEC method conditions for proteins

Proteins provide a unique challenge in robustness testing and liquid chromatographic methods development, as the profile is typically less resolved than with small molecules. As with their small molecule counterparts, traditional elements of validation are required for regulatory approval. Due to the complexity of the chromatographic profile, the elements of robustness can be particularly challenging to evaluate for methods involving antibodies.

Robustness evaluation of the size exclusion chromatography (SEC) and hydrophobic interaction chromatography (HIC) methods can be efficiently evaluated using a design of experiments (DOE) platform. Ganorkar et al. remind us that DOE is a series of experimental designs using statistical techniques for planning, conducting, analysing and interpreting data from experiments.¹

Liquid chromatography (LC) methods to characterise antibody-drug conjugate (ADC) formulations typically involve SEC and HIC. SEC methods are used to characterise the higher and lower molecular weight of the ADC. The effect of column temperature, flow rate, and need for a salt or organic modifier is typically addressed in method robustness studies.

Download this article to discover the challenges, opportunities and optimisations needed to execute design of experiment studies ADCs.