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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.