Abstract
Antigen Used for Immunization
2×105 CFU of F. tularensis holarctica strain LVS, followed by an outer membrane-enriched LVS preparation. 1
Method of Immunization
Intradermal priming of a BALB/cJ mouse with LVS, followed 48 days later by intraperitoneal boosting with the membrane-enriched prep adjuvanted with CpG ODN 1826 (TCC ATG ACG TTC CTG ACG TT), an oligodeoxynucleotide containing unmethylated CpG dinucleotides with fully phosphorothioate backbone, 3.75 days before euthanasia.
Parental Cell Line Used for Fusion
Sp2/0-Ag14.
Selection and Cloning Procedure
Screening of hybridoma supernatants by ELISA and Western blot analysis on membrane-enriched LVS prep and LVS lysate, respectively. Cloning by limiting dilution and selection of wells with single clones and ELISA reactivity to LVS vortexate.
Heavy and Light Chains of Immunoglobulin
Mouse γ2a, κ.
Specificity
Internal epitope of F. tularensis O-antigen, as determined by ladder pattern in Western blot analysis on F. tularensis LPS, which shows decreasing reactivity with decreasing LPS chain length and inhibition of its binding to LPS in ELISA by internal-binding but not terminal-binding MAbs specific for F. tularensis O-antigen.
Specific Antigen Identified
F. tularensis O-antigen of LPS and capsule.