Characterization of Shigella flexneri-specific murine monoclonal antibodies by chemically defined glycoconjugates

J Immunol. 1986 Oct 1;137(7):2361-6.

Abstract

Chemically defined glycoconjugates are demonstrated to have considerable potential for selecting hybridoma antibodies directed toward O-antigenic determinants, especially when used in combination with a panel of well-characterized LPS molecules. Monoclonal antibodies specific for the Shigella flexneri O-antigens of serogroup 5b, variants X and Y, were generated after immunization of BALB/c mice with killed bacterial cells, and active hybrids were selected on the basis of ELISA performed with the purified serotype-specific LPS antigen. Subsequent screening with a variety of glycoconjugates, derived from synthetic oligosaccharides and larger structures obtained by phage Sf6/endo-rhamnosidase hydrolysis of purified LPS established a detailed profile of binding characteristics for Shigella flexneri variant Y-specific antibodies. Together with the results of precipitin analysis and heavy chain isotyping experiments, a limited number of antibodies were selected as candidates for detailed studies of the antibody combining site.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology*
  • Antibody Affinity
  • Antibody Specificity
  • Antigens, Bacterial / immunology
  • Enzyme-Linked Immunosorbent Assay
  • Mice
  • Polysaccharides, Bacterial / chemical synthesis
  • Polysaccharides, Bacterial / immunology*
  • Serotyping
  • Shigella flexneri / classification
  • Shigella flexneri / immunology*

Substances

  • Antibodies, Monoclonal
  • Antigens, Bacterial
  • Polysaccharides, Bacterial