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Open Access Highly Accessed Research article

Identification of immunogenic proteins and generation of antibodies against Salmonella Typhimurium using phage display

Torsten Meyer1, Thomas Schirrmann1, André Frenzel1, Sebastian Miethe1, Janin Stratmann-Selke23, Gerald F Gerlach2, Katrin Strutzberg-Minder2, Stefan Dübel1 and Michael Hust1*

Author Affiliations

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

2 IVD GmbH Heisterbergallee 12, 30453 Hannover, Germany

3 Present address: vaxxinova GmbH diagnostics, Johann-Krane-Weg 42, 48149 Münster, Germany

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BMC Biotechnology 2012, 12:29  doi:10.1186/1472-6750-12-29

Published: 15 June 2012

Abstract

Background

Solely in Europoe, Salmonella Typhimurium causes more than 100,000 infections per year. Improved detection of livestock colonised with S. Typhimurium is necessary to prevent foodborne diseases. Currently, commercially available ELISA assays are based on a mixture of O-antigens (LPS) or total cell lysate of Salmonella and are hampered by cross-reaction. The identification of novel immunogenic proteins would be useful to develop ELISA based diagnostic assays with a higher specificity.

Results

A phage display library of the entire Salmonella Typhimurium genome was constructed and 47 immunogenic oligopeptides were identified using a pool of convalescent sera from pigs infected with Salmonella Typhimurium. The corresponding complete genes of seven of the identified oligopeptids were cloned. Five of them were produced in E. coli. The immunogenic character of these antigens was validated with sera from pigs infeced with S. Tyhimurium and control sera from non-infected animals. Finally, human antibody fragments (scFv) against these five antigens were selected using antibody phage display and characterised.

Conclusion

In this work, we identified novel immunogenic proteins of Salmonella Typhimurium and generated antibody fragments against these antigens completely based on phage display. Five immunogenic proteins were validated using a panel of positive and negative sera for prospective applications in diagnostics of Salmonela Typhimurium.