Innovative Aufreinigungsstrategien auf Membranadsorberbasis für rekombinante Zytokine

Abstract

Downstream processing accounts for substantial cost in the production of cytokines. To overcome the limitations of commonly used packed bed chromatography columns, innovative cation exchange membrane adsorbers as a basis for rapid bioseparation are presented. Here, the transport of solutes to the binding sites takes place predominantly by convection, thereby reducing both process time and eluate volume. Moreover, scale up of membrane chromatography is simple due to its modular format. While membrane chromatography in direct capture mode integrates clarification and protein capture into one single unit operation and thus reduces the number of stages in the downstream process. Additionally, we introduce new weak and strong cation exchange membrane adsorbers combining mixed mode ligand chemistry and an optimized membrane platform for the purification of cytokines. The results show that the ligand chemistry as well as the membrane matrix can have impact on the selectivity, binding and the recovery of the cytokine.The membrane-based bioseparation technology made large scale purification of high-value proteins possible. Implementation of a functional upscaling process was required for economic feasibility. We introduce three model proteins to apply alternative purification strategies using membrane chromatography. Due to their unique structural motive, purification is rather challenging. High protein aggregation and renaturation steps accounts for complexity of the isolation process. Another aspect is the implementation of an animal component free process. Here, membrane adsorber strategies substitute conventional approaches using heparin chromatography. This plays in important role for the certification of the product regarding purity.