Monoclonal antibodies are the antibodies derived from a single B-cell clone and are specific for single epitope. In 1975, George Kohler and Ceasar Milstein developed a method for preparing monoclonal antibodies. They were produced by Hybridoma Technology. A hybrid cell called Hybridoma, is made by the fusion of normal activated antibody producing B-cell with a myeloma cell. Plama B-cell are capable of producing antibodies and Myeloma cells have immortal growth properties. This Hybridoma has properties of both of these cells.
- Mix spleen cells from a mouse that has been immunized with the desired antigen, with Myeloma cells.
- Use a Fusogen to facilitate fusion of plasma membrane like PEG.
- Cell Fusion Mixture is transferred to HAT selection Medium. HAT contains Hypoxanthine Aminopterin and Thymidine.
- Unfused Myeloma cells cannot grow because they lack HGPRT .
- Unfused Normal Spleen cells cannot grown indefinitely because of their limited life span.
- Hybridoma cells are able to grow indefinitely on HAT medium because spleen cells provide HGPRT and myeloma cells provide Immortality.
- Monoclonal antibody is used in diagnostic and therapeutic agents in clinical medicine.
- They are also used for detecting pregnancy, pathogenic micro-organisms, to measure the concentration of various drugs in the blood.