2. Contents
◦ Introduction
◦ Monoclonal Antibodies production
◦ Hybridoma Technique
◦ Applications of Monoclonal Antibodies (Biomedical
Research/Medicine)
◦ Side effects of monoclonal antibodies
◦ References
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3. Introduction
◦ Monoclonal antibodies are clones of your body's antibodies that are made in a
laboratory, meant to stimulate your immune system.
◦ Monoclonal antibodies can have monovalent affinity, binding only to the
same epitope.
◦ Antibodies are produced by B cells (specialized WBCs). When an antigen
comes into contact with a B cell, it causes the B cell to divide and clone.
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4. Monoclonal Abs Polyclonal Abs
◦ Monoclonal antibodies are very specific, and
only bind to a particular antigen structure. They
therefore can only target a specific chemical or
specific cells in the body.
◦ Epitope Specific
◦ Expensive to develop
◦ Slow to produce (approx. 6 months)
◦ Polyclonal antibodies are still specific to a
particular antigen, but they can recognise
different variations of the same antigen.
◦ Not Epitope Specific
◦ Inexpensive to develop
◦ Quick to produce (approx. 3 months)
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6. How are Monoclonal Antibodies are
produced?
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◦ Hybridoma technology is one of the most common methods used to produce
monoclonal antibodies. In this process, antibody-producing B lymphocytes are
isolated from mice after immunizing the mice with specific antigen and are
fused with immortal myeloma cell lines to form hybrid cells, called hybridoma
cell lines.
◦ Georges Kohler and Cesar Milstein in 1975 invented the hybridoma
technology, for which they received the Nobel Prize in 1984 in physiology and
medicine.
7. Procedure
◦ Insert the antigen into a mouse. The monoclonal antibody you want to produce
will have a specific antigen. This antigen is inserted into a mouse to initiate an
immune response in the mouse.
◦ The mouse immune response occurs. The mouse’s immune response takes place
and antibodies are created against the supposed pathogen. You may want to go
back to the tutorial on the immune system to remind you how antibodies are
produced.
◦ B Lymphocytes are taken from the mouse. The mouse currently has lots of B
lymphocytes making our desired antibody. The spleen produces lymphocytes,
so we can harvest (take) some B lymphocytes from the mouse spleen.
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8. ◦ The lymphocytes are combined with myeloma cells. We can’t just grow the B
lymphocytes, because they divide very slowly. Therefore we can combine these
slow-growing B lymphocytes with fast growing tumor cells called myeloma
cells. This forms fast-growing hybridoma cells.
◦ HAT media is added to remove unfused & fused cells. So only Hybridoma cells
remain at the end. This is called selection. These cells are allowed to grow then
individual colonies are picked up and cultured. Specificity of the antibody is
tested by adding antigen, specific to that antibody. (ELISA)
◦ Many antibodies are produced in the bio reactor. The hybridoma cells are then
cloned and used to produce many cells that all make the same antibody. The
antibodies can be collected and purified.
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9. Clinical Research Programs, Gujarat University
The myeloma cells
(fused/unfused)
die because they are HGPRT
negative. They cannot use the
salvage pathway The
normal B-cells/fused divide for
few days then
eventually die because they are
mortal.
1.
2.
3.
4.
5.
6.
7.
8.
9.
11. Applications of Monoclonal Antibodies
◦ Biomedical Research- ELISA, Western blot, Immunofluorescence, Chromatin
Immunoprecipitation.
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12. ◦ Immunotheraphy/ Antibodies used as Medicine: eg. Rituximab, Alemtuzumab
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14. What monoclonal antibodies are used for
COVID-19?
• Bebtelovimab is authorized to treat outpatients only and not patients who have
been admitted to the hospital with serious symptoms of COVID-19 and require
oxygen therapy.
◦ Evusheld (tixagevimab/cilgavimab) is to prevent or protect someone from
becoming ill before they are exposed to COVID-19. It is not used to treat
COVID symptoms or prevent someone from illness after being exposed to the
virus. Evusheld is not a substitute for COVID vaccination.
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15. What are the side effects of monoclonal antibodies?
Common side effects of monoclonal antibodies include:
◦ Allergic reactions
◦ Chills
◦ Weakness
◦ Diarrhea
◦ Nausea
◦ Vomiting
◦ Rash
◦ Itching
◦ High blood glucose levels
◦ Cough
◦ Constipation Clinical Research Programs, Gujarat University
16. ◦ Serious infections are more likely to occur when monoclonal antibodies are
combined with other drugs that suppress the immune system (for example,
steroids).
◦ Monoclonal antibodies may interfere with the effectiveness of vaccines. Live
vaccines, including attenuated vaccines, should not be used while patients
are being treated with monoclonal antibodies. Patients should complete all
recommended immunizations prior to receiving monoclonal antibodies.
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