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Star Republic: Guide for Biologists

Monoclonal antibody production schedule

Phase I: IMMUNIZATION
1-2 months
  • Immunization of 3-5 Balb/c female mice.
  • Antibody capture ELISA to determine best two mice to be used for cell fusion.
  • Phase II: FUSION
    2-3 weeks
  • Fusion of spleen cells from the two best mice.
  • Selection and expansion of fused clones.
  • Determination of positive hybridomas by antibody capture ELISA assay.
  • Phase III: EXPANSION AND SUBCLONING
    1.5-3 months
  • Isolation of antibody-secreting colonies.
  • Further testing of positive clones.
  • Subcloning by limiting dilution.
  • Determination of isotypes.
  • Expansion of monoclonal cell lines.
  • Collection of monoclone antibody containing supernatant.