Ion exchange chromatography (IEX) is an established technique for aggregate removal in downstream processing of therapeutic proteins. In polishing processing flow through mode is known for high productivity, low buffer consumption and homogeneous product concentration profile. Membrane adsorbers (MA) offer significant advantageous over resin based systems when used for contaminant and/or aggregate removal. The main benefits of MA compared to chromatographic resin are higher mass transfer rates and thus elevated productivity, as well as ease-of-use due to the pre-packed and often also single use nature of the MA devices. While batch isotherms usually display stationary parameters, the presented flow through (FT) approach can determine dynamic parameters like displacement effects. Here we describe a high throughput screening (HTS) robotic technique for parameter determination and optimization for FT aggregate removal by MA. Two HTS methods are introduced for aggregate removal of a recombinant CHO cell fermented and post protein A clarified immunoglobulin G feed. The classical bind and elute (BE) mode for process parameter screening investigates conductivity and pH-value influence onto the separation. BE mode is optimized for low protein mass consumption by connecting salt steps to obtain selectivity and binding capacity with one loaded MA. For dynamic investigations the advanced bind and elute (ABE) mode is introduced. In ABE mode the elution step CD is used for the next loading step to investigate the initial salt concentration influence on dynamic effects. The eight parallel down-scale HTS set up is used for characterization of cation exchange Sartobind® Q and anion exchange Sartobind® S MA. The screening includes different pH-values and salt concentrations. BE and ABE HTS mode deliver detailed information’s in IgG aggregate removal and possible processing parameter and steps.

Aggregate removal in polishing mAb process step with membrane chromatography by determining binding capacity and displacement effects in a HTS robotic set-up

2019 Spring National Meeting

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