Cancer is a disease that arises from abnormal growth of the body’s cells that turn into cancer cells. The disease can be triggered by multifactors such as genetics, carcinogens (agents that can encourage the formation of cancer cells), viruses, and unhealthy lifestyles.
The body has an immune system that is responsible for foreign attacks of microbes or from transformed cells. One such form of defense is played by an important molecule called an antibody.
Therefore, the use of monoclonal antibodies, a biologically based drug that can recognize cancer antigens specific, for cancer therapy is a revolutionary step in the last few decades in the world.
Monoclonal antibodies are one type of treatment that triggers (encourages) the body’s immune system to fight a disease (immunotherapy). These monoclonal antibodies are an artificial immune or immune protein specifically designed to mark specific cancer cells.
Thus, it can kill cancer cells without damaging and or destroying the healthy cells that are nearby. Some types of cancer that can use monoclonal antibodies as a therapeutic option are breast cancer cells, skin cancer, blood cancer and colorectal cancer.
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Treatment of Cancer Types and Options
Monoclonal Antibodies Types
Monoclonal antibodies have 4 types, namely:
- Murine, purely obtained from mice can cause human anti mouse antibodies (HAMA) to name its ending “momab” (ibritumomab).
- Chimeric, combined Fc human antibodies and Fab monoclonal antibodies mice, the sling name “ximab” (rituximab).
- Humanized, only a small fraction of Fab mouse antibodies combine with human antibodies (95-98%) the sling name is “zumab” (trastuzumab).
- Fully human, the whole human antibody, the sling names “mumab” (adalimumab).
Basiliximab is a monoclonal antibody that prevents the proliferation of T lymphocytes, used for acute rejection prophylaxis in allogeneic kidney transplants. Administered with cyclosporine and corticosteroid immunosuppressants, their use should be limited by experts.
Rituximab is a monoclonal antibody that causes B lymphocyte lysis, used for the treatment of advanced follicular lymphoma that has been resistant to chemotherapy and for the diffusion of large B cell non-Hodgkin lymphoma combined with other chemotherapy.
Rituximab is combined with other chemotherapy used for the treatment of advanced follicular lymphoma that has not received treatment. Complete facilities for resuscitation should be prepared and like other cytotoxic treatments should be carried out under the supervision of an oncologist.
Monoclonal Antibodies Advantages
Some advantages of using immunotherapy in eradicating cancer cells are:
- It can kill cancer cells more effectively when compared to radiation therapy and chemotherapy treatment
- It can help the performance of other therapies
- It has relatively few side effects compared to others
- It can recognize cancer cell types so as to reduce recurrence cases
Monoclonal Antibodies Production
Kőhler and Milstein explain how to isolate and develop large quantities of specific pure monoclonal antibodies obtained from a mixture of immune response antibodies.
Mice that have been immunized with special antigens into the bone marrow will produce B lymphocyte cells that have a limited lifesizing period in the culture, this can be overcome by combining with perennial B lymphocyte cells (myeloma).
The result of a heterogeneous mixture of hybridoma cells selected hybridoma that has 2 capabilities that can produce special antibodies and it can grow in cultures. This hybridoma is reproduced according to its individual clones and each clone produces only one permanent and stable type of monoclonal antibody.
Hybridoma derived from one lymphocyte will produce antibodies that will recognize one type of antigen. These antibodies known as monoclonal antibodies.
The process of making monoclonal antibodies through 5 stages, namely
Immunization of mice and selection of donor mice in the development of hybridoma cells.
Mice are immunized with certain antigens to produce the desired antibodies. Mice were killed if the antibody was sufficiently reached in serum and then the spleen was used as a source of cells to be combined with myeloma cells.
Screening the production of serum antibody.
Antibodies in the blood of mice were assessed after several weeks of immunization. Titer serum antibodies are determined by various techniques such as enzyme linked immunosorbent assay (ELISA) and flow cytometry.
Cell fusion can be done if the antibody titer is already high if the titer is still low then it should be done booster until a strong response is achieved. The production of hybridoma cells in vitro is taken from the spleen of mice were killed.
Preparation of myeloma cells.
Myeloma cells obtained from perennial lymphocyte tumors cannot grow if the deficiency of hypoxanthine guanine phosphoribosyl transferase (HGPRT) and normal spleen cells is limited in life’s end. The antibodies from spleen cells that have a limited life end provide HGPRT and are then combined with myeloma cells, which life lasts so that a hybridoma can grow indefinitely.
Myeloma cells are perennial cells cultured with 8 azaguanine sensitive to the medium selection of hypoxanthine aminopterin thymidine (HAT). One week before cell fusion, myeloma cells are cultured in 8 azaguanine. Cells must have the high life ability and be able to grow fast. Cell fusion uses the HAT medium to survive in a culture.
Fusion of myeloma cells with spleen immune cells
One spleen cell is combined with the prepared myeloma cell. This fusion is resolved through centrifugation of spleen cells and myeloma cells in polyethylene glycol, a substance that can combine cell membranes.
Cells that successfully experience fusion can grow on a special medium. The cells were then distributed into a place containing food, obtained from the peritoneal fluid of mice. The cell’s food source provides a growth factor for hybridoma cell growth.
Further development of hybridoma cell cloning
A small group of hybridoma cells can be developed in tissue cultures by means of antigen bond selection or developed through mouse ascites method. Cloning by limiting dilution will ensure a clone is successful.
Hybridoma cultures can be sustained in vitro in culture tubes (10-60 ug/ml) and in vivo in mice, living in rat ascites. The concentration of antibodies in serum and other bodily fluids is 1-10 ug/ml.
