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Infection and Immunity, May 2002, p. 2487-2491, Vol. 70, No. 5
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.5.2487-2491.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Analysis of Factors Affecting Surface Expression and Immunogenicity of Recombinant Proteins Expressed by Gram-Positive Commensal Vectors

Tové C. Bolken,1 Christine A. Franke,1 Kevin F. Jones,1,2 Richard H. Bell,1 Ryan M. Swanson,1 David S. King,1 Vincent A. Fischetti,3 and Dennis E. Hruby1,2*

SIGA Technologies Inc., Corvallis, Oregon 97333,1 Department of Microbiology, Oregon State University, Corvallis, Oregon 97330,2 Laboratory of Bacterial Pathogenesis and Immunology, The Rockefeller University, New York, New York 100213

Received 24 October 2001/ Returned for modification 16 January 2002/ Accepted 28 January 2002

Several key protein structural attributes were altered in an effort to optimize expression and immunogenicity of a foreign protein (M protein from Streptococcus pyogenes) exposed on the surface of Streptococcus gordonii commensal bacterial vectors: (i) a shorter N-terminal region, (ii) the addition of a 94-amino-acid spacer, and (iii) the addition of extra C-repeat regions (CRR) from the M6 protein. A decrease in the amount of cell surface M6 was observed upon deletion of 10 or more amino acid residues at the N terminus. On the other hand, reactivity of monoclonal antibody to surface M6 increased with the addition of the spacer adjacent to the proline- and glycine-rich region, and an increase in epitope dosage was obtained by adding another CRR immediately downstream of the original CRR. The results obtained should facilitate the design of improved vaccine candidates using this antigen delivery technology.

* Corresponding author. Mailing address: SIGA Technologies Inc., 4575 S.W. Research Way, Suite 230, Corvallis, OR 97333. Phone: (541) 753-2000. Fax: (541) 753-9999. E-mail: dhruby{at}sgph.com.

Editor: E. I. Tuomanen

Infection and Immunity, May 2002, p. 2487-2491, Vol. 70, No. 5
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.5.2487-2491.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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