Advertisement

Expression of scFv in L-Form Bacteria

Part of the Springer Lab Manuals book series (SLM)

Abstract

The production of heterologous protein, including recombinant antibody derivatives like scFv, in the periplasmic space of E. coli is accomplished by correct signal sequence processing, formation of disulfide bridges and can therefore lead to soluble functional recombinant product. But the yield of functional, soluble recombinant protein from E. coli is often drastically limited by growth inhibition due to a toxic effect of the product and protein aggregation. In accordance with other investigators concerning scFv expression in E. coli we found that in the periplasm only a constant threshold concentration of soluble recombinant protein can be reached. Moreover this threshold concentration is dependent on the individual V-regions and can not be significantly influenced by promoter strength and culture conditions (Rippmann et al., 1998).

Keywords

Proteus Mirabilis Recombinant Antibody scFv Antibody Promoter Strength Soluble Recombinant Protein 
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.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Dübel S, Breitling F, Klewinghaus I, and Little M (1992) Regulated secretion and purification of recombinant antibodies in E. coli. Cell. Biophys. 21: 69–79PubMedGoogle Scholar
  2. Dübel S, Breitling F, Fuchs P, Braunagel M, Klewinghaus I, and Little M (1993) A family of vectors for surface display and production of antibodies. Gene. 128: 97–101PubMedCrossRefGoogle Scholar
  3. Gumpert J, Cron H, Plapp R, Niersbach H, and Hoischen C (1996) Synthesis and secretion of recombinant penicillin G acylase in bacterial L-forms. J. Basic Microbiol. 36: 89–98PubMedCrossRefGoogle Scholar
  4. Gumpert, J. and Hoischen, C. (1998). Use of cell wall-less bacteria (L-forms) for efficient expression and secretion of heterologous gene products. Curr. Opin. Biotechnol. 9, 506–509.PubMedCrossRefGoogle Scholar
  5. Gumpert J and Taubeneck U (1983) Characteristic properties and biological significance of stable protoplast type L-forms. Experientia Suppl. 46: 227–241PubMedGoogle Scholar
  6. Kujau, M.J., Hoischen, C., Riesenberg, D., and Gumpert, J. (1998). Expression and secretion of functional miniantibodies McPC603scFvDhlx in cell-wall-less L-form strains of Proteus mirabilis and Escherichia coli: a comparison of the synthesis capacities of L-form strains with an E. coli producer strain. Appl. Microbiol. Biotechnol. 49, 51–58.PubMedCrossRefGoogle Scholar
  7. Rippmann JF, Klein M, Hoischen C, Brocks B, Rettig J, Gumpert J, Pfizenmaier K, Mattes R, and Moosmayer D (1998) Procaryotic expression of scFv antibodies: secretion in L-form cells of Proteus mirabilis leads to active product and overcomes limitations of periplasmic expression in Escherichia coli. Applied and Environmental Microbiology 64:4862–4869PubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2001

Authors and Affiliations

  1. 1.Institute of Cell Biology and ImmunologyUniversity of StuttgartStuttgartGermany
  2. 2.Institute of Industrial GeneticsUniversity of StuttgartStuttgartGermany
  3. 3.Boehringer Ingelheim Pharma KGGermany

Personalised recommendations