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© Research
Publication :

High Yield Production Process for Shigella Outer Membrane Particles

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in - 06 Jun 2012

Berlanda-Scorza, F., A.M. Colucci, L. Maggiore, S. Sanzone, I. Pesce, I. Ferlenghi, N. Norais, V. Di Cioccio, A. Saul, and C. Gerke

Link to Pubmed [PMID] – 22701551

PLoS One. 2012;7(6):e35616

Gram-negative bacteria naturally shed particles that consist of outer membrane lipids, outer membrane proteins, and soluble periplasmic components. These particles have been proposed for use as vaccines but the yield has been problematic. We developed a high yielding production process of genetically derived outer membrane particles from the human pathogen Shigella sonnei. Yields of approximately 100 milligrams of membrane-associated proteins per liter of fermentation were obtained from cultures of S. sonnei ΔtolR ΔgalU at optical densities of 30–45 in a 5 L fermenter. Proteomic analysis of the purified particles showed the preparation to primarily contain predicted outer membrane and periplasmic proteins. These were highly immunogenic in mice. The production of these outer membrane particles from high density cultivation of bacteria supports the feasibility of scaling up this approach as an affordable manufacturing process. Furthermore, we demonstrate the feasibility of using this process with other genetic manipulations e.g. abolition of O antigen synthesis and modification of the lipopolysaccharide structure in order to modify the immunogenicity or reactogenicity of the particles. This work provides the basis for a large scale manufacturing process of Generalized Modules of Membrane Antigens (GMMA) for production of vaccines from Gram-negative bacteria

http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0035616