We demonstrate the application of scanning electron microscopy (SEM), finite bath uptake experiments, confocal laser scanning microscopy (CLSM) and small scale column studies to understanding chromatography fouling. Two systems were compared; a hydrophobic interaction capture step for virus-like particles and an anion exchange polishing step following a protein A affinity capture. In both systems resin samples taken for SEM and finite bath uptake experiments indicate that as fouling proceeds with successive batch cycles significant blockage of the pores at the resin surface occurs, thereby decreasing protein uptake rate. However morphological differences between the two systems were seen. Further study, conducted using CLSM to allow temporal and spatial measurement within the resins, was performed using clean, partially fouled and fully fouled resin samples. Packed within a miniaturized flow cell the results indicated that the foulant severely decreasing the protein uptake rate, while a minimal decrease in saturation capacity was found.

Understanding chromatography fouling in vaccine and therapeutic protein manufacture

243rd National Meeting (2012)

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