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Logo of bmcbiotBioMed Centralsearchsubmit a manuscriptregisterthis articleBMC Biotechnology
BMC Biotechnol. 2012; 12: 29.
Published online Jun 15, 2012. doi:  10.1186/1472-6750-12-29
PMCID: PMC3423037
Identification of immunogenic proteins and generation of antibodies against Salmonella Typhimurium using phage display
Torsten Meyer,1 Thomas Schirrmann,1 André Frenzel,1 Sebastian Miethe,1 Janin Stratmann-Selke,2,3 Gerald F Gerlach,2 Katrin Strutzberg-Minder,2 Stefan Dübel,1 and Michael Hustcorresponding author1
1Technische Universität Braunschweig, Institut für Biochemie, Biotechnologie und Bioinformatik, Abteilung Biotechnologie, Spielmannstr.7, 38106 Braunschweig, Germany
2IVD GmbH Heisterbergallee 12, 30453 Hannover, Germany
3Present address: vaxxinova GmbH diagnostics, Johann-Krane-Weg 42, 48149 Münster, Germany
corresponding authorCorresponding author.
Torsten Meyer: torsten.meyer/at/; Thomas Schirrmann: th.schirrmann/at/; André Frenzel: andre.frenzel/at/; Sebastian Miethe: s.miethe/at/; Janin Stratmann-Selke: janin/at/; Gerald F Gerlach: gfgerlach/at/; Katrin Strutzberg-Minder: strutzberg/at/; Stefan Dübel: s.duebel/at/; Michael Hust: m.hust/at/
Received January 23, 2012; Accepted May 25, 2012.
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.
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.
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.
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