P33: Loss Avoidance:  Realizing efficiency and yield improvement targets in the primary clarification step of high cell density fermentations

Monday, November 7, 2011
Capri Ballroom (Marriott Marco Island)
D. Gregory Cybulski, Sales, Celeros Separations, Foxboro, MA
Recent advances in the understanding industrialized yeasts such as Pichia pastoris, with its ability for rapid fermentation and high cell growth, have expanded their prevalence as an accepted antibody producing platform.  In the process of purifying antibodies from high cell density fermentations, the primary clarification step is the principal opportunity to improve upon substantial losses associated with historical yield and efficiency expectations.   In this analysis we evaluate an advanced centrifugal separation technology as a source for loss avoidance in processes where post-fermentation dilution rates and multi-pass centrifuge protocols are the norm.  The APD centrifuge was challenged to clarify a P. pastoris broth having a cell density of 150 OD600 at 45% wet weight.  As an examination of clarification efficiency, OD600 measurements of centrate samples were monitored to ensure centrifuge optimization.  Requiring no dilution and under a single-pass centrifugation strategy, the APD effectively processed the batch while maintaining centrate OD levels within desired specifications.  Reaffirming the advantage of low liquid entrainment in this step, the paste was weighed in support of mass balance calculations.  Observations from these trials indicate elevated yields, and suggest that high value improvement targets remain attainable in the rapidly evolving field of antibody manufacturing.
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