Optimizing the expression of a monoclonal antibody fragment under the transcriptional control of the Escherichia coli lac promoter

Can J Microbiol. 2000 Jun;46(6):532-41.

Abstract

The expression of a monoclonal antibody Fab fragment in Escherichia coli strain RB791/pComb3, induced with either lactose or isopropyl-beta-D-thiogalactoside (IPTG), was compared to determine if lactose might provide an inexpensive alternative to induction with IPTG. Induction of Fab expression imposed a metabolic load on the recombinant cells, resulting in lower final cell yields compared to the non-induced controls. An IPTG concentration of 0.05 mM was sufficient to achieve maximal expression of soluble Fab protein when inducing in the early-, mid-, or late-log phases of batch cultures grown using either glucose or glycerol as a carbon source. The largest overall yield of Fab fragments when using 0.05 mM IPTG was achieved by increasing the final yield of cells through glycerol feeding following induction in late-log phase. Lactose was as effective as IPTG for inducing Fab expression in E. coli RB791/pComb3. The greatest overall level of Fab expression was found when cells grown on glycerol were induced with 2 g/L lactose in late-log phase. Since the cost of 0.05 mM of IPTG is significantly greater than the cost of 2 g/L lactose, lactose provides an inexpensive alternative to IPTG for inducing the expression of Fab fragments, and possibly other recombinant proteins, from the E. coli lac promoter.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / biosynthesis*
  • Blotting, Western
  • Culture Media
  • Escherichia coli / genetics*
  • Escherichia coli / growth & development
  • Escherichia coli / metabolism
  • Glucose / metabolism
  • Glycerol / metabolism
  • Immunoglobulin Fab Fragments / biosynthesis*
  • Isopropyl Thiogalactoside / pharmacology
  • Lac Operon*
  • Lactose / pharmacology
  • Promoter Regions, Genetic
  • Recombinant Proteins / biosynthesis
  • Transcriptional Activation
  • beta-Galactosidase / metabolism

Substances

  • Antibodies, Monoclonal
  • Culture Media
  • Immunoglobulin Fab Fragments
  • Recombinant Proteins
  • Isopropyl Thiogalactoside
  • beta-Galactosidase
  • Glucose
  • Lactose
  • Glycerol