Evaluating the impact of high Pluronic® F68 concentrations on antibody producing CHO cell lines.
Pluronic® F68 (P-F68) is an important component of chemically-defined cell culture medium because it protects cells from hydrodynamic and bubble-induced shear in the bioreactor. While P-F68 is typically used in cell culture medium at a concentration of 1 g/L (0.1%), higher concentrations can offer additional shear protection and have also been shown to be beneficial during cryopreservation. Recent industry experience with variability in P-F68-associated shear-protection has opened up the possibility of elevated P-F68 concentrations in cell culture media, a topic that has not been previously explored in the context of industrial cell culture processes. Recognizing this gap, we first evaluated the effect of 1-5 g/L P-F68 concentrations in shake flask cultures over ten 3-day passages for cell lines A and B. Increase in terminal cell density and cell size was seen over time at higher P-F68 concentrations but protein productivity was not impacted. Results from this preliminary screening study suggested no adverse impact of high P-F68 concentrations. Subsequently fed-batch bioreactor experiments were conducted at 1 and 5 g/L P-F68 concentrations with both cell lines where cell growth, viability, metabolism, and product quality were examined under process conditions reflective of a commercial process. Results from these bioreactor experiments confirmed findings from the preliminary screen and also indicated no impact of elevated P-F68 concentration on product quality. If additional shear protection is desired, either due to raw material variability, cell line sensitivity, or a high-shear cell culture process, our results suggest this can be accomplished by elevating the P-F68 concentration in the cell culture medium without impacting cell culture performance and product quality.
Cell Science & Technology, Process & Product Development, Amgen Inc. One Amgen Center Drive, Thousand Oaks, 91320, California.
Tharmala
T
Tharmalingam
Chetan T
CT
Goudar
D000818:Animals; D000906:Antibodies / Q000235:genetics / Q000378:metabolism*; D019149:Bioreactors; D016466:CHO Cells / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D049109:Cell Proliferation / Q000187:drug effects; D048429:Cell Size / Q000187:drug effects; D003412:Cricetulus; D020442:Poloxamer / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D000906:Antibodies; D011994:Recombinant Proteins; D020442:Poloxamer
Pluronic® F68; bioreactor; cell culture; product quality; shear protection