This work examined the impact of using a single-pass tangential flow filtration (SPTFF) as an in-line concentration step to concentrate cell culture harvest process streams prior to downstream purification. Harvest process streams can create facility bottlenecks due to limited hold tank capacity and extended load times for Protein A affinity chromatography that can be overcome by SPTFF. However, SPTFF can be susceptible to membrane fouling especially in the presence of complex mixtures of proteinaceous particulates, host cell proteins (HCP), DNA, and media components found in harvest process streams. Prior to deciding operating parameters for SPTFF, cell culture harvest from three different monoclonal antibodies were processed through a chromatographic clarifier (CC), and recirculated under total recycle using 30 kDa and 100 kDa regenerated cellulose ultrafiltration membrane. The sieving coefficients of HCP and residual DNA were measured. Insights into membrane fouling on exposure to harvest were obtained for these antibodies. The sieving data was used to simulate impurity clearance for SPTFF system using mass balance models. This was verified by actual SPTFF experiments. The addition of a chromatographic clarifier followed by a single-pass concentration reduced the protein-A loading time and offered better impurity reduction post affinity chromatography.

Investigation of single pass tangential flow filtration as an inline concentration step during cell culture harvest

251st National Meeting (2016)

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