Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
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Generation of Isotype Switch Variants form Hybridoma cells Producing anti-Streptococcus penumoniae 6B Polysaccharide Antibody

  • Kim, Jihye (Faculty of Biological Sciences, Chonbuk National University, Chonju) ;
  • Eunja Ryu (Faculty of Biological Sciences, Chonbuk National University, Chonju) ;
  • Park, Moon-Kook (Faculty of Biological Sciences, Chonbuk National University, Chonju)
  • Published : 1999.09.01
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Abstract

hybridoma cells producing IgM anti-pneuococcal 6B polysaccharide antibodies were induced to switch to IgG-producing cells in vitro by treating with acridine orange. Treating 0.5 $\mu\textrm{g}$/ml of acridine orange for 24 hours generated maximal number of variant cells. The maximal isotype switch frequency was 3${\times}$10-5, which is about 30-fold higher than the frequency of spontaneous switching. Resulting IgG-producing variants were enriched by sib selection and ELISA spot assay. Two IgG3-producing variant cells were finally cloned by limiting dilution. The variant cells produced similar amounts of antibodies as their parental cells did. The two switched antibodies had similar reactivity to pneumococcal 6B polysaccharide. When compared to their parental IgM antibodies, the switched IgG3 than that of IgM antibody. The antibodies will be useful as essential tools for comparative study of the role of heavy chain isotypes in protection against Streptococcus pneumoniae.

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