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Infection and Immunity, August 2003, p. 4633-4641, Vol. 71, No. 8
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.8.4633-4641.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Functional Selection of Vaccine Candidate Peptides from Staphylococcus aureus Whole-Genome Expression Libraries In Vitro

Thomas Weichhart,1 Markus Horky,1 Johannes Söllner,1 Susanne Gangl,1 Tamàs Henics,1 Eszter Nagy,1 Andreas Meinke,1 Alexander von Gabain,1 Claire M. Fraser,2 Steve R. Gill,2 Martin Hafner,1 and Uwe von Ahsen1*

Intercell AG, 1030 Vienna, Austria,1 The Institute for Genomic Research, Rockville, Maryland 208502

Received 24 February 2003/ Returned for modification 24 April 2003/ Accepted 14 May 2003

An in vitro protein selection method, ribosome display, has been applied to comprehensively identify and map the immunologically relevant proteins of the human pathogen Staphylococcus aureus. A library built up from genomic fragments of the virulent S. aureus COL strain (methicillin-resistant S. aureus) allowed us to screen all possible encoded peptides for immunoreactivity. As selective agents, human sera exhibiting a high antibody titer and opsonic activity against S. aureus were used, since these antibodies indicate the in vivo expression and immunoreactivity of the corresponding proteins. Identified clones cluster in distinct regions of 75 genes, most of them classifiable as secreted or surface-localized proteins, including previously identified virulence factors. In addition, 14 putative novel short open reading frames were identified and their immunoreactivity and in vivo mRNA expression were confirmed, underscoring the annotation-independent, true genomic nature of our approach. Evidence is provided that a large fraction of the identified peptides cannot be expressed in an in vivo-based surface display system. Thus, in vitro protein selection, not biased by the context of living entities, allows screening of genomic expression libraries with a large number of different ligands simultaneously. It is a powerful approach for fingerprinting the repertoire of immune reactive proteins serving as target candidates for active and passive vaccination against pathogens.

* Corresponding author. Mailing address: Intercell AG, Campus Vienna Biocenter 6, 1030 Vienna, Austria. Phone: 43 1 20620-220. Fax: 43 1 20620-800. E-mail: uahsen{at}intercell.com.

Editor: D. L. Burns

Infection and Immunity, August 2003, p. 4633-4641, Vol. 71, No. 8
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.8.4633-4641.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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