Increasing cell culture densities and productivities during therapeutic protein (MAbs) production are placing a larger burden on downstream clarification and purification operations due to higher product and impurity levels. Controlled flocculation and/or precipitation of mammalian cell culture suspensions by chemical means using either acids or polymers has been used as an alternative technology to enhance the clarification throughput and downstream filtration operations. While flocculation is quite effective in agglomerating cell debris and adventitious (host cell proteins and DNA), the resulting suspension is generally not easily separable by ordinary direct filtration methods. Traditionally, centrifugation and a combination of filtration techniques (tangential-flow filtration and depth filtration) have been widely used for clarifying these pretreated cell culture broths. However, the increased cellular debris present in these complex feed streams can prematurely foul the membrane and commercially available depth filters, adversely impacting their capacity and throughput. In this work, EMD Millipore has developed novel filter designs which when coupled with cell culture suspension optimization result in improved primary and secondary clarification for flocculated high cell density mammalian cell cultures systems feeds. The design of the filter media and the interactions between the flocculation method and the filter will be discussed. Potential benefits will be discussed, including more robust processing without a centrifuge and the potential to completely replace centrifugation technologies with direct harvest filtration techniques.

Clarification of recombinant proteins from high cell density mammalian cell cultures systems using new improved depth filters

243rd National Meeting (2012)

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