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Controlling Apoptosis to Optimize Yields of Proteins from Mammalian Cells

  • Matthew P. ZustiakEmail author
  • Haimanti Dorai
  • Michael J. Betenbaugh
  • Tina M. Sauerwald
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 801)

Abstract

Apoptosis is the foremost method of cell death in bioreactors and can be caused by nutrient limitation, toxin accumulation, and growth factor withdrawal. By delaying the onset of this form of programmed cell death, one can achieve longer sustained viabilities in culture, thereby increasing product yield. Described here is a genetic-based, step-by-step method to generate an apoptosis-resistant cell line. This cell line, then, can be used as a platform for biotherapeutic protein production. The key steps include antiapoptotic transgene selection and transfection followed by clonal isolation and screening. With the proper screening methods, one can obtain a robust cell line that resists the harsh conditions of late-stage and/or high-density culture.

Key words

Apoptosis Transfection Selection Screening Flow cytometry bcl-XL CHO Monoclonal antibody Mammalian cell culture 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Matthew P. Zustiak
    • 1
    Email author
  • Haimanti Dorai
    • 2
  • Michael J. Betenbaugh
    • 1
  • Tina M. Sauerwald
    • 2
  1. 1.Department of Chemical and Biomolecular EngineeringJohns Hopkins UniversityBaltimoreUSA
  2. 2.Gene Expression, Centocor, Inc.RadnorUSA

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