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Identification of immunogenic polypeptides from a Mycoplasma hyopneumoniae genome library by phage display

  • Applied Genetics and Molecular Biotechnology
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Abstract

The identification of immunogenic polypeptides of pathogens is helpful for the development of diagnostic assays and therapeutic applications like vaccines. Routinely, these proteins are identified by two-dimensional polyacrylamide gel electrophoresis and Western blot using convalescent serum, followed by mass spectrometry. This technology, however, is limited, because low or differentially expressed proteins, e.g. dependent on pathogen–host interaction, cannot be identified. In this work, we developed and improved a M13 genomic phage display-based method for the selection of immunogenic polypeptides of Mycoplasma hyopneumoniae, a pathogen causing porcine enzootic pneumonia. The fragmented genome of M. hyopneumoniae was cloned into a phage display vector, and the genomic library was packaged using the helperphage Hyperphage to enrich open reading frames (ORFs). Afterwards, the phage display library was screened by panning using convalescent serum. The analysis of individual phage clones resulted in the identification of five genes encoding immunogenic proteins, only two of which had been previously identified and described as immunogenic. This M13 genomic phage display, directly combining ORF enrichment and the presentation of the corresponding polypeptide on the phage surface, complements proteome-based methods for the identification of immunogenic polypeptides and is particularly well suited for the use in mycoplasma species.

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Acknowledgements

We would like to thank Stefan Dübel for corrections and discussion on the manuscript and Saskia Helmsing for technical assistance.

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  1. Jonas Kügler & Simone Nieswandt

    Present address: Helmholtz Zentrum für Infektionsforschung, Inhoffenstr. 7, 38124, Braunschweig, Germany

Authors and Affiliations

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

    Jonas Kügler, Simone Nieswandt, Thomas Schirrmann & Michael Hust

  2. Institut für Mikrobiologie, Stiftung Tierärztliche Hochschule Hannover, Bischofsholer Damm 15, 30173, Hannover, Germany

    Gerald F. Gerlach & Jochen Meens

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  1. Jonas Kügler
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Correspondence to Michael Hust.

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Kügler, J., Nieswandt, S., Gerlach, G.F. et al. Identification of immunogenic polypeptides from a Mycoplasma hyopneumoniae genome library by phage display. Appl Microbiol Biotechnol 80, 447–458 (2008). https://doi.org/10.1007/s00253-008-1576-1

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  • DOI: https://doi.org/10.1007/s00253-008-1576-1

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