Monoclonal antibodies are laboratory-produced molecules engineered to serve as substitute antibodies that can restore, enhance, or mimic the immune system’s attack on cells. They are designed to bind to specific targets found on cancer cells. These are identical immune cells that are all clones of a unique parent cell and target one specific antigen. Created using hybridoma technology, which involves fusing an antibody-producing B cell with a myeloma (cancer) cell, resulting in a hybrid cell (hybridoma) that can produce large quantities of a specific antibody.

Steps in Monoclonal Antibody Production

1.Antigen Preparation: Identify and purify the antigen that the monoclonal antibody will target. This could be a protein or other molecule associated with a disease.
2.Immunization of Mice: Inject the antigen into mice to stimulate an immune response.
Perform multiple injections over a few weeks to boost the immune response, leading to the production of B cells that generate the desired antibodies.
3.Cell Fusion (Hybridoma Formation):Harvest spleen cells from the immunized mice, which include the B cells producing antibodies against the antigen.
Fuse these spleen cells with myeloma cells (a type of cancer cell that can grow indefinitely) using a fusion agent like polyethylene glycol (PEG). This creates hybrid cells, or hybridomas, that can produce the specific antibody and divide endlessly.
4.Selection of Hybridomas: Grow the hybridomas in a selective medium, typically Hypoxanthine-Aminopterin-Thymidine (HAT) medium, which allows only the fused cells to survive.
Screen the surviving hybridomas to identify those producing the desired antibody. This is done using assays such as enzyme-linked immunosorbent assay (ELISA).
5.Cloning:Isolate single hybridoma cells and allow them to proliferate to ensure that each cell line is producing a monoclonal antibody.
Perform limiting dilution cloning to obtain pure hybridoma cell lines.
6.Production and Purification: Expand the selected hybridoma cell lines in culture.
Harvest the monoclonal antibodies from the culture medium.
Purify the antibodies using techniques like protein A/G affinity chromatography, ion exchange chromatography, or size-exclusion chromatography.

Test the purified monoclonal antibodies to confirm their specificity, affinity, and functionality. This can involve various biochemical and biophysical assays.

Uses:

• Cancer Treatment: Target cancer cells specifically, minimizing damage to normal cells.
• Autoimmune Diseases: Can modulate the immune system in diseases like rheumatoid arthritis or multiple sclerosis.
• Infectious Diseases: Used in the treatment of conditions like COVID-19.
• Diagnostics: Utilized in various diagnostic tests to detect pathogens.