Hybridoma technology is the classical method of monoclonal antibody production and an effective way of producing antibodies on a large scale. It involves the fusion of antibody producing B-cells with long-lived myeloma cells.
1. After immunization against the antigen of choice, splenocytes are isolated from the host.
2. Splenocytes are fused with myeloma cells lacking the HGPRT gene.
3. Cells are grown in HAT medium to select for successfully fused hybridomas.
4. Hybridomas are separated to single cell level and screened by e.g. ELISA.
5. Positive clones are identified and re-cloned.
6. The hybridoma colony is grown in supplemented culture medium until fully established.
7. Cells are transferred to bioreactors for large scale production.
8. Antibodies are harvested and purified.