Use of sulfated-cellulose membrane adsorbers to intensify purification of cell culturederived influenza A and B viruses

New generation of cell culture-based vaccines enables faster response to pandemic outbreaks and helps coping with the increasing demand for seasonal vaccines. Besides efficient upstream processing technologies, appropriate economic and robust downstream processing is key to consistently achieve high yields. Chromatography membranes have been extensively evaluated to capture viruses at laboratory scale, for influenza, adenovirus or virus like particles. They have shown great potential to intensify processes thanks to a high binding capacity, high flow-rate and ease of use and are now commonly used in new generation of vaccine processes. We present here Sartobind SC, a novel sulfated cellulose membrane adsorber, for the capture of Influenza. In a recent study, the binding capacity and the purification performance of two bead-based resins and Sartobind SC was compared for three influenza virus strains (H1N1, H3N2 and B) produced in MDCK suspension cells in a chemically defined medium. The dynamic binding capacity for the sulfated cellulose membrane adsorbers was consistently higher than for the resins (8 to 22-fold). Overall, recovery of virus varied between 66% and 81%. Total protein and DNA removal were\u3e74% and\u3e96%, respectively. Due to the higher operating flow rate and binding capacity, the productivity with the membrane adsorbers was on average 25-times higher than with the resins. This purification platform based on sulfated cellulose can intensify processes and therefore reduce the cost of influenza purification. This ligand is also a promising candidate for other viruses such as vaccinia, RSV, HSV or measles

Scalable, Robust and Highly Productive Novel Convecdiff Membrane Platform for mAb Capture

The recombinant monoclonal antibody capture step represents the current bottleneck in downstream processing. Protein A resins are diffusion-limited chromatography materials which require low flow rates to achieve a binding capacity above 30 g L−1 with the result of low productivity. Here, we present a novel chromatography membrane combining superior binding capacities with high flow rates for high productivity while achieving comparable product quality as state-of-the-art protein A resins. Further, we demonstrate full scalability of this convecdiff technology with experimental data demonstrating suitability for bioprocessing at different scales. This technology results in more than 10-fold higher productivity compared to Protein A resins, which is maintained during scale up. We demonstrate the influence of residence times, feed titers and the cleaning regime on productivity and indicate optimal utilization of the convecdiff membrane based on feed titer availability. The underlying high productivity and short cycle times of this material enable the purification of monoclonal antibodies with 10-times less chromatography material used per batch and utilization of the membrane within one batch. Provided in disposable consumables, this novel technology will remove column handling in bioprocesses and resin re-use over multiple batches

Development of a Single-Step Antibody–Drug Conjugate Purification Process with Membrane Chromatography

Membrane chromatography is routinely used to remove host cell proteins, viral particles, and aggregates during antibody downstream processing. The application of membrane chromatography to the field of antibody-drug conjugates (ADCs) has been applied in a limited capacity and in only specialized scenarios. Here, we utilized the characteristics of the membrane adsorbers, Sartobind® S and Phenyl, for aggregate and payload clearance while polishing the ADC in a single chromatographic run. The Sartobind® S membrane was used in the removal of excess payload, while the Sartobind® Phenyl was used to polish the ADC by clearance of unwanted drug-to-antibody ratio (DAR) species and aggregates. The Sartobind® S membrane reproducibly achieved log-fold clearance of free payload with a 10 membrane-volume wash. Application of the Sartobind® Phenyl decreased aggregates and higher DAR species while increasing DAR homogeneity. The Sartobind® S and Phenyl membranes were placed in tandem to simplify the process in a single chromatographic run. With the optimized binding, washing, and elution conditions, the tandem membrane approach was performed in a shorter timescale with minimum solvent consumption and high yield. The application of the tandem membrane chromatography system presents a novel and efficient purification scheme that can be realized during ADC manufacturing