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Kinetic Analysis of Recombinant BHK Roller Bottle Cultures

  • Frank Deer
  • Jennifer Mahoney
  • Abner Correia
  • Kevin Stafford
Conference paper
Part of the ESACT Proceedings book series (ESACT, volume 5)

Abstract

A BHK cell line that secretes recombinant porcine Factor VIII is being used in a roller bottle based manufacturing process to supply clinical product for Phase III trials. A chemically-defined medium is used to feed confluent cultures in iterative fashion over a 5 day period, during which the recombinant product is harvested on a daily basis. In order to better understand process capability, a study was performed to more closely examine the kinetics of recombinant protein production over each 24 h iterative batch period. The results showed that the majority of recombinant product was secreted within the first 18 h of the incubation period, with no additional product being made over the final 6 h interval. Spent media analysis revealed that certain amino acids were notably depleted by the end of each day. Another study was performed to evaluate whether recombinant protein production could be boosted during the final 12 h of the batch period. The amino acids which were rapidly depleted during the initial study were added back to production cultures halfway through the batch period on each production day. This strategy resulted in a 23% increase in rpFVIII yield by the end of the production process.

Keywords

Recombinant Protein Production Recombinant Product Roller Bottle Batch Period Recombinant Porcine 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Frank Deer
    • 1
  • Jennifer Mahoney
    • 1
  • Abner Correia
    • 2
  • Kevin Stafford
    • 3
  1. 1.Ipsen BiomeasureMilfordUSA
  2. 2.New England Controls, Inc.FoxboroughUSA
  3. 3.Inspiration BiopharmaceuticalsLaguna NiguelUSA

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