The document describes hybridoma technology, which involves fusing antibody-producing B cells from immunized mice with myeloma cells to produce hybridomas that can indefinitely produce monoclonal antibodies of a single specificity. The key steps are immunizing mice, fusing B cells with myeloma cells, selecting antibody-producing hybridomas through cloning or limiting dilution, screening for the desired antibody, and expanding the clones. Hybridoma technology revolutionized immunology and provides a powerful method for producing large quantities of highly specific monoclonal antibodies.
2. INTRODUCTION OF HYBRIDOMA
TECHNOLOGY
• The discovery of hybridoma technology was made by César Milstein and Georges
Köhler in 1975, for which they were awarded the Nobel Prize in Physiology or
Medicine in 1984.
• Hybridoma technology is a powerful method for producing large quantities of
highly specific monoclonal antibodies. The hybridomas are capable of producing a
single type of antibody, known as a monoclonal antibody, which can recognize
and bind to a specific antigen.
• The development of hybridoma technology revolutionized the field of
immunology and has become an important tool for biomedical research, clinical
diagnostics, and therapeutics.
3. PROCEDURE OF HYBRIDOMA TECHNOLOGY
The process of hybridoma
technology involves several
steps:
1. IMMUNIZATION
2. FUSION
3. SELECTION
4. SCREENING OF PRODUCTS
5. CLONING
4. An animal(mouse), typically a
mouse, is immunized with a specific
antigen. The antigen is injected into
the animal's bloodstream, which
triggers an immune response and the
production of antibodies.
IMMUNIZATION
5. FUSION
The B lymphocytes are then fused with
myeloma cells, which are cancerous
cells that can divide indefinitely. The
fusion process is typically induced by
treating the cells with a chemical or
electric shock.
6. SELECTION
Selection of a single antibody producing
hybrid cells is very important. This is
possible if the hybrids are isolated and grown
individually. The suspension of hybrid cells
is so diluted that the individual liquors
contain on an average one cell each. These
cells, when grown in a regular culture
medium, produce the desired antibody.
7. SCREENING OF PRODUCTS
Screening of the product in hybridoma
technology is a crucial step to ensure that the
resulting monoclonal antibody is specific and
of high quality
ELISA and RIA are commonly used of
screening of product.
ELISA SCREENING METHOD
8. CLONING
1.Limiting dilution: The hybridoma cells are
first diluted in a culture medium, such that each
well of a culture plate receives only one cell.
This process is called limiting dilution.
The selected hybridoma cells are then
cloned to ensure that all the cells in a
particular clone produce the same
monoclonal antibody. This is achieved
through limiting dilution or other
techniques.
9. Advantages of hybridoma technology:
1.Specificity: Hybridoma technology produces monoclonal antibodies that are highly specific
and can recognize a single epitope on the antigen of interest.
2.Consistency: Cloning of hybridoma cells ensures that all the cells in a particular clone
produce the same monoclonal antibody, providing consistency in the production of the
antibody.
3.Scalability: Hybridoma technology can produce large quantities of monoclonal antibodies,
making it suitable for industrial-scale production.
4.Diversity: Hybridoma technology can be used to produce monoclonal antibodies against a
wide range of antigens, including proteins, peptides, and small molecules.
5.Versatility: Monoclonal antibodies produced by hybridoma technology have a wide range
of applications, including research, diagnostics, and therapeutics.
10. Disadvantages of hybridoma technology:
1. Animal use: Hybridoma technology requires the use of animals, typically mice, for the
production of monoclonal antibodies, which raises ethical concerns.
2. Time-consuming: The production of monoclonal antibodies by hybridoma technology
can be time-consuming, as it involves multiple steps, including immunization, cell
fusion, screening, cloning, and expansion.
3. Cost: The cost of producing monoclonal antibodies by hybridoma technology can be
high, particularly for large-scale production.
4. Limited antigen types: Hybridoma technology is limited to producing monoclonal
antibodies against antigens that can induce an immune response in animals.
5. Limited production of human antibodies: Hybridoma technology can produce mouse
monoclonal antibodies, but it is more challenging to produce human monoclonal
antibodies using this method.
