Open AccessAnalysis of Factors Affecting Surface Expression and Immunogenicity of Recombinant Proteins Expressed by Gram-Positive Commensal Vectors
Author(s) -
Tove′ C. Bolken,
Christine A. Franke,
Kevin F. Jones,
Richard H. Bell,
Richard A. Swanson,
David S. King,
Vincent A. Fischetti,
Dennis E. Hruby
Publication year - 2002
Publication title -
infection and immunity
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.508
H-Index - 220
eISSN - 1070-6313
pISSN - 0019-9567
DOI - 10.1128/iai.70.5.2487-2491.2002
Subject(s) - immunogenicity , streptococcus gordonii , biology , recombinant dna , epitope , streptococcus pyogenes , monoclonal antibody , antigen , amino acid , microbiology and biotechnology , antibody , streptococcus , biochemistry , bacteria , gene , immunology , genetics , staphylococcus aureus
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.