Background & Novelty: One of the raw materials used in mammalian cell culture media responsible for loss in performance especially in large scale CHO fed-batch processes is Poloxamer 188 (Peng H. ; 2014). Poloxamer is a water-soluble triblock copolymer of polyethyleneoxyde (PEO) and polypropylene oxide (PPO) and is part of an important class of non-ionic surfactants containing a hydrophobic center (PPO) and two hydrophilic tails (PEO). The function remains still unclear (Gigout A.; 2008) but often causes lot-to-lot inconsistencies published and described independently by various biopharmaceutical companies. The function of the 8400 Dalton block polymer is described differently e.g. as shear protecting agent (Sowana D.; 2002) and Poloxamer prevented loss of cell viability in a gas sparged or bubble-free environment (Tharmalingam T.; 2008) and thus makes correlation of the malfunction directly related to properties of this molecule difficult. To correlate the function of the molecule with one physico-chemical property is not successful up to now as the reason for loss in viable cell density in bioreactors is may be not mono-causal. The objective of this project was to generate Poloxamer 188 with consistent quality by the development of a new cell culture grade material to assure consistency and reliable cell culture performance verified by the established small scale cell-culture assay. Experimental Procedure: First we have established a cell culture assay to analyze the lot-to-lot inconsistency in a small scale cell culture assay. Besides the cell culture analytics a broad panel of physico-chemical methods (like IR, NMR, HPLC, GC and others) were applied to discriminate good performing Poloxamer lots from suspect or non-performing lots. Afterwards we produced and processed the polymer with a unique set-up of technologies the functionality of Poloxamer was verified using a generic CHO-S cell assay (modified from Peng et al., 2014) Results and discussion: The usage of Poloxamer is crucial for the performance in large scale CHO-processes. Therefore functional assays and a production procedure to assure consistent quality are key assets to support this biopharmaceutical industry need. We will show how this was addressed from a combined approach in analytics and production to launch scalable quantities of cell culture grade Poloxamer.

Poloxamer: Steps towards cell culture media consistency

251st National Meeting (2016)

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