Poster Presentation BacPath 13: Molecular Analysis of Bacterial Pathogens Conference 2015

Production of a heterologous protein vaccine candidate through an autotransporter based platform (#102)

Irene Beriotto 1 2 , Yanina Sevastsyanovich 2 , Giacomo Romagnoli 3 , Silvana Savino 3 , Christiane Gerke 4 , Allan J. Saul 1 , Francesca Micoli 1 , Adam F. Cunningham 2 , Calman A. MacLennan 5 , Ian Henderson 2
  1. Sclavo Behring Vaccines Institute for Global Health S. r. L (a GSK Company), Siena, Italy
  2. Institute of Microbiology and Infection, University of Birmingham, Birmingham, United Kingdom
  3. Novartis Vaccines and Diagnostics (a GSK Company), Siena, Italy
  4. Direction du Développement / Grant Office, Institut Pasteur, Paris, France
  5. The Jenner Institute, University of Oxford, Oxford, United Kingdom

Recently, we developed a method for the selective extracellular accumulation of recombinant proteins using an autotransporter secretion pathway. This technology involves engineering of the Pet autotransporter protein to allow secretion of a protein of interest into the culture medium. In this work we report how the platform can be exploited for the production of an ETEC protein vaccine candidate. This production system offers a valid alternative to other traditional methods for production of “difficult” proteins with comparable results in terms of purity and protein conformational stability thus having a simplified and cost effective downstream process. The antigen production using the Pet autotransporter system has been scaled for the first time up to a fermentation level, obtaining up to 29 mg of antigen produced per litre of culture supernatant. Taken together these findings suggest the suitability of the system for further industrial applications.