Following up on our post introducing antibody-based experiments, we wanted to describe how antibodies are made in large volume: First, an animal is immunized with the protein (or peptide) of interest; Second, the spleen is dissected and the plasma cells producing the antibodies of interested are isolated; Finally, these plasma cells are fused with myeloma cells to “immortalize” them into a hybridoma (an antibody-producing cell-line that can be cultured indefinitely).
There are multiple methods to culture hybridomas and harvest large quantities of antibody from the supernatant. My favorite method is to use: CELLine1000 Bioreactors are able to grow hybridomas to very high densities by virtue of (1)a 1 Liter media reservoir which enables continuous nutrient in-flow and waste out-flow and (2) an oxygen permeable base which allows continuous oxygen inflow and carbon dioxide out-flow. With this setup I have obtained 15 mg of antibody per week per bioreactor which I could rapidly purify by commercial Protein G spin columns.
- Howard, G.C.; Kaser, M.R. Making and Using Antibodies: A Practical Handbook CRC Press 2006
- Alberts, B. Molecular Biology of the Cell 5th Ed. Garland Science 2008
- Murphy, K. Janeway’s Immunobiology 8th Ed. Garland Science 2012
This work by Eugene Douglass and Chad Miller is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.