Monoclonal antibodies are set of antibodies which are target specific against a single source of antigen. They are obtained from single set of cells and hence named as monoclonal antibodies. They are extensively used in medical field and are in huge demand due to their application like in treatment of diseases of some foreign substance like infections, cancer cells or for diagnosis etc. They are also used in research of biology and related medical fields.
1. First an animal like a mouse, rabbit etc. is immunized with suitable antigen which corresponds to desired monoclonal antibody we are interested in. This is done by injecting antigens which are previously emulsified with some adjuvants like Freund’s adjuvant.
2. This leads to enhnaced production of desired antibodies in the mouse body. The immunisation is done for few weeks till the antibody concentration in the mouse blood reaches a desired level.
3. After several weeks, the blood test is done to see antibody titre using suitable techniques like ELISA or flow cytometry etc.and are obatined from mouse spleen.
4. Then the obtained monoclonal antibodies are fused with a cancer cell (i.e. myeloma not just tumor) as they are immortal and can divide indefinitely.
5. So the fused cell grow indefinitely to produce huge number of monoclonal antibodies..The bunch of fused cells is called hybridoma.
6. These formed tumor cells are injected into mouse peritoneal cavity or grown in vitro by tissue culture technique.
8. On injection into mouse peritoneal cavity, the cell grow rapidly and release them in the form of luid into the abdomen which appear as bulged fluid bags (ascites).
9. These monoclonal antibodies from ascites are extracted to harvest the formed antibodies.
Another method is growing these hybridoma cells invitro but is not easy and also is expensive than the mouse method. But sometimes the mouse may experience pain, irritation due to ascites. Then they go for invitro cell culture technique.