Unit

UNIT 5.10 Production of Recombinant Proteins in Mammalian Cells

  1. Su Chen,
  2. David Gray,
  3. Johnny Ma,
  4. Shyamsundar Subramanian

Published Online: 1 MAY 2001

DOI: 10.1002/0471140864.ps0510s12

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Chen, S., Gray, D., Ma, J. and Subramanian, S. 2001. Production of Recombinant Proteins in Mammalian Cells. Current Protocols in Protein Science. 12:5.10:5.10.1–5.10.41.

Author Information

  1. Chiron Corporation, Emeryville, California

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JUN 1998

Abstract

The best strategy for consistent production of larger quantities of pure protein is stable expression. Popular hosts for stable expression are Chinese hamster ovary (CHO) cells, baby hamster kidney (BHK-21) cells, myeloma cells, and the transformed kidney cell line 293. Protocols for stable production in CHO cells are described in this unit. Typical methods for transfection using commercially available plasmid expression vectors are described, along with methods to select for stable expression and methods for amplifying the expression level in the transfected cell. Following this, procedures are presented for efficient cell growth to obtain significant amounts of protein product. Support protocols describe freezing of cells, determination of growth rates, determination of specific productivity of cells, preparing samples for assay, and setting up a 10-day shaker-flask growth curve.