Membrane chromatography has been explored as a viable alternative to packed-bed column chromatography for the purification of therapeutic proteins. Purification via membrane chromatography offers several advantages, including higher productivity and reduced buffer usage. As is the case with packed-bed chromatography, the best approach for the development of a membrane chromatography purification step is to utilize high-throughput screening in order to reduce development times and material requirements. This research focused on the development of a new, high-throughput screening technique for use in screening multiple membrane chromatography conditions for monoclonal antibody purification. The developed screen utilizes a specialized 96 well plate that contains 19µl of embedded membrane in each well. This setup allows for the screening of multiple different membranes at once, and four mixed-mode cation exchange membranes and one cation exchange membrane were evaluated on the plate as part of this research. The plate was used first to map out partitioning coefficients for the different membranes using equilibrium binding capacity screens, and the results from this initial screen were used to map feasible process conditions. A secondary screen was performed in an attempt to mimic the continuous loading in a larger scale purification run. A discreet loading scheme allowed for the fractionation of the flowthrough, and therefore yields and purities could be measured across the entire loading range. Results from this screen were compared to purification runs on larger-scale (0.6 ml) membrane disks, and the results demonstrated that the screen produced similar yield and aggregate clearance trends across the entire range of tested loadings. Therefore, this research demonstrated the effective use of this screen in a development strategy that enabled the optimization of multiple conditions in less time and with less material compared to laboratory-scale purification runs.

High-throughput approach to developing and optimizing mixed-mode membrane chromatography for protein purification

2019 Spring National Meeting

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