Antibody Purification from Cell Culture Supernatant Using New Polymeric Multimodal Membranes

This poster will describe our latest research efforts on devising improved methods to capture and purify antibodies from cell culture supernatant using overload and elute chromatography with new multimodal membranes (MMMs). The membranes incorporate functional groups that provide orthogonal modes of interactions, which dramatically increases the range of ionic strength for operation of these membranes relative to conventional ion-exchange and hydrophobic interaction chromatography media, and makes them attractive candidates to face emerging challenges in downstream processing of biologics...

Protein binding experiments with IgG show the MMM has a high equilibrium binding capacity (180 mg IgG/ml) and excellent salt tolerance. It maintains binding capacities in excess of 100 mg IgG/ml at salt concentrations that are typical for elution buffers used in multi-stage chromatographic bioseparation processes. Protein dynamic binding capacities in excess of 50 mg IgG/ml were determined from breakthrough curve analysis.

In one study, the MMMs were used as the stationary phase following a Protein A purification step in the purification human IgG1 from Chinese hamster ovary (CHO) cell culture supernatant. The MMM stationary phase greatly enhanced load productivity compared to resin-based ion-exchange media. Further, no buffer exchange step was required between the Protein A and MMM chromatography purification steps.

In a second study, the MMMs were used as the stationary phase for the primary capture step, in place of Protein A resin chromatography. By loading the same column volumes of feed solution, the MMMs demonstrated a high selectivity for capturing human IgG from CHO cell culture supernatant. A high quality protein product was obtained from the MMM chromatography purification step.

This work shows that the new MMM chromatography operation can effectively decrease the number of purification steps and shorten the purification time and/or increase load productivity in downstream processing of biologics. It further shows that the new MMMs may be suitable for capture step purification of non-mAb proteins or when Protein A cannot be used.