<br/>Beta-Glucans are polysaccharides of D-glucose monomers linked by (1-3) beta-glycosidic bonds and are found to have a potential immunogenicity risk, labeling them as process contaminants. A common source of beta-glucans is from the cellulose found in traditional depth filter media. While beta-glucans are introduced into the process from cell culture raw materials and the depth filters during primary clarification, they can be removed by the subsequent bind and elute affinity chromatography step. However, historical data has shown that, unlike bind and elute mode, flowthrough mode chromatography, both ionic exchange and hydrophobic, cannot remove beta-glucan. Thus, the primary focus of this study was to control the beta-glucans that leached out from the depth filtration step following viral inactivation and neutralization and prior to the flowthrough mode polishing steps. An optimized depth filter flush strategy using pH 10 Sodium Carbonate buffer was developed to control beta-glucan levels while maintaining flowthrough operation of the subsequent polishing steps. Different loading conditions for the depth filtration and subsequent chromatography steps were evaluated to determine the robustness of the optimized flush strategy. Carry through runs have resulted in greater than 2 fold reduction in beta-glucan levels for the optimized wash.

Control of beta-glucan levels in downstream processing by improved depth filtration wash

2019 Spring National Meeting

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