Technical Material

Poloxamer 188 (P188, Pluronic F-68) is a raw material, which is routinely used in cell culture media to protect cells from hydrodynamic forces and plays an important role in the overall process performance (1,2). Previous studies suggest that if additional shear protection is required (for example, due to cell-line sensitivity or high-shear culture process), then this can be accomplished by increasing Poloxamer concentrations (up to 5 g/L), without impacting cell culture performance and product quality (2). 

Here, we present the effect of various concentrations (0-2 g/L) of Poloxamer 188 (‘Kolliphor®, Sigma-Aldrich) on both cell culture performance and productivity. These results show that, although small increase in maximum viable cell density (VCD) can be observed for higher concentration of poloxamer, there is no significant effect on IgG production. Taken together, we recommend the addition of around 1 g/L Poloxamer for optimal culture performance.

The graphs below shows the effect of Poloxamer 188 concentration on viable cell density, viability and IgG titer, in antibody-producing CHO-DG44 cell line:

Note: Previously, considerable lot-to-lot variations of Poloxamer 188 made it difficult to include as raw material in powder cell- culture media (3, 4). However, recent improvements now allow consistent performance, and Ajinomoto Genexine’s latest CHO - cell media, BASAL4P, now includes Kolliphor® P 188 Bio, a high purity poloxamer designed specifically to reduce shear stress during biologics manufacturing.

References:

1) Guzniczak E., et al. (2018). Impact of poloxamer 188 (Pluronic F-68) additive on cell mechanical properties, quantification by real-time deformability cytometry. Biomicrofluidics 12(4):044118.

2) Tharmalingam, T., Goudar, C. (2015). Evaluating the Impact of High Pluronic (R) F68 Concentrations on Antibody Producing CHO Cell Lines. Biotechnology and bioengineering. 112(4):832-7.

3) Peng, H., et al. (2014). Development of small scale cell culture models for screening poloxamer 188 lot‐to‐lot variation. Biotechnol Progress, 30(6): 1411-8.

4) Narayanappa , A.T., et al. (2019). A rapid cell-based assay for determining poloxamer quality in CHO suspension cell culture. BioTechniques 2019 67:3, 98-109.