Characterization of monoclonal antibodies to terminal and internal O-antigen epitopes of Francisella tularensis lipopolysaccharide

Hybridoma (Larchmt). 2011 Feb;30(1):19-28. doi: 10.1089/hyb.2010.0083.


The lipopolysaccharide (LPS) of Francisella tularensis (Ft), the Gram negative bacterium that causes tularemia, has been shown to be a main protective antigen in mice and humans; we have previously demonstrated that murine anti-Ft LPS IgG2a monoclonal antibodies (MAbs) can protect mice against otherwise lethal intranasal infection with the Ft live vaccine strain (LVS). Here we show that four IgG2a anti-LPS MAbs are specific for the O-polysaccharide (O-antigen [OAg]) of Ft LPS. But whereas three of the MAbs bind to immunodominant repeating internal epitopes, one binds to a unique terminal epitope of Ft OAg. This was deduced from its even binding to both long and short chains of the LPS ladder in Western blots, its rapid decrease in ELISA binding to decreasing solid-phase LPS concentrations, its inability to compete for LPS binding with a representative of the other three MAbs, and its inability to immunoprecipitate OAg despite its superior agglutination titer. Biacore analysis showed the end-binding MAb to have higher bivalent avidity for Ft OAg than the internal-binding MAbs and provided an immunogenicity explanation for the predominance of internal-binding anti-Ft OAg MAbs. These findings demonstrate that non-overlapping epitopes can be targeted by antibodies to Ft OAg, which may inform the design of vaccines and immunotherapies against tularemia.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Antibodies, Monoclonal / genetics
  • Antibodies, Monoclonal / immunology*
  • Antibody Specificity / immunology*
  • Binding, Competitive / immunology
  • Blotting, Western
  • Enzyme-Linked Immunosorbent Assay
  • Epitopes / genetics
  • Francisella tularensis / immunology*
  • Immunoglobulin G / genetics
  • Immunoglobulin G / immunology*
  • Immunoprecipitation
  • Lipopolysaccharides / immunology*
  • O Antigens / genetics
  • O Antigens / immunology*


  • Antibodies, Monoclonal
  • Epitopes
  • Immunoglobulin G
  • Lipopolysaccharides
  • O Antigens