Raising the Antibodies - Monoclonal antibodies for a specific enzyme or protein are raised using traditional hybridoma technology. The enzyme is first extracted and partially purified using most of the conventional steps, such as extraction, ammonium sulfate precipitation and gel column filtration.

The preparation is diluted, and emulsified with 'Freunds complete adjuvant'.Then it is injected intramuscularly into mice, to raise antibodies. Generally several doses at intervals of a few days are injected to boost the antibody production.

The spleen cells from the mice are collected and fused with myeloma (tumor) cell lines derived from a rat. The resultant hybridoma cells are screened for their capacity to produce antibodies and secrete them out. The hybridoma ,cells can be thawed and recultured for the production of antibodies.