Research

Production of recombinant antibody fragments in Bacillus megaterium

Eva Jordan¹ , Michael Hust¹ , Andreas Roth² , Rebekka Biedendieck² , Thomas Schirrmann¹ , Dieter Jahn² and Stefan Dübel¹

¹  Technische Universität Braunschweig, Institut für Biochemie und Biotechnologie, Abteilung Biotechnologie, Spielmannstr. 7, 38106 Braunschweig, Germany

²  Technische Universität Braunschweig, Institut für Mikrobiologie, Spielmannstr. 7, 38106 Braunschweig, Germany

author email corresponding author email

Microbial Cell Factories 2007, 6:2doi:10.1186/1475-2859-6-2

Published:	15 January 2007

Abstract

Background

Recombinant antibodies are essential reagents for research, diagnostics and therapy. The well established production host Escherichia coli relies on the secretion into the periplasmic space for antibody synthesis. Due to the outer membrane of Gram-negative bacteria, only a fraction of this material reaches the medium. Recently, the Gram-positive bacterium Bacillus megaterium was shown to efficiently secrete recombinant proteins into the growth medium. Here we evaluated B. megaterium for the recombinant production of antibody fragments.

Results

The lysozyme specific single chain Fv (scFv) fragment D1.3 was succesfully produced using B. megaterium. The impact of culture medium composition, gene expression time and culture temperatures on the production of functional scFv protein was systematically analyzed. A production and secretion at 41°C for 24 h using TB medium was optimal for this individual scFv. Interestingly, these parameters were very different to the optimal conditions for the expression of other proteins in B. megaterium. Per L culture supernatant, more than 400 μg of recombinant His₆-tagged antibody fragment were purified by one step affinity chromatography. The material produced by B. megaterium showed an increased specific activity compared to material produced in E. coli.

Conclusion

High yields of functional scFv antibody fragments can be produced and secreted into the culture medium by B. megaterium, making this production system a reasonable alternative to E. coli.