Generation of Murine Monoclonal Antibodies by Hybridoma Technology

The overall goal of this procedure is to generate antibody producing hybridoma cell lines for the biotechnological production of monoclonal antibodies. This method can be used to generate specific binders for the detection and visualization of biomolecules and for the development of effective detection systems, like ELISA, flow cytometry, or magnetic cell sorting. The main advantage of this technique is that it facilitates the generation of monoclonal antibody producing cell lines with a predefined specificity and a continuous availability.

To prepare the myeloma cells, thaw a vial of murine SP2/0 cells in warm water. Then transfer the cells into 10 mL of culture medium and collect them by centrifugation. Re-suspend the pellet in 10 mL of fresh culture medium and seed the cells into a 25 square centimeter flask for incubation at 37 degrees celsius, and 5%CO2, preserving aliquots of the cells for longterm storage after the appropriate number of expansions.