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.
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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.
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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.
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