Hybridoma technology

Introduction

When two non-identical cells are brought into close contact and their membranes are allowed to fuse, the resulting product contains both nuclei. The fusion can occur between cells of the same species or of different species. A cell with two or more dissimilar nuclei is called a heterokaryon and their fusion results in a single nucleus. The fusion product is called a hybrid. And the technology is Hybridoma technology.

The hybrids lose some of the chromosomes but individual properties are retained by the parental cells. In 1975, Kohler and Milstein fused antibody-producing mouse spleen cells with mouse myeloma cells. This experiment has led to increased use of somatic cell hybridization.

Significance in biomedical research

The techniques used for the production of monoclonal antibodies and the technology, known as hybridoma technology. This technology has proved capable of producing rodent antibodies of predetermined specificity to a wide variety of different antigens.

Hybridoma technology allows the production of hybrid cell lines from B cells that secrete a single, monoclonal antibody with one binding specificity.

Hence, they can thus be produced in unlimited quantities.

Role in the production of monoclonal antibodies

Hybridoma technology has played a key role in the production of monoclonal antibodies. Some of the factors include:
High specificity: Hybridoma technology promotes the production of highly specific and consistent monoclonal antibodies that are essential for medical research purposes.
Unlimited quantity of identical antibodies: With the advancement in hybridoma technology, it became possible to produce large quantities of identical monoclonal antibodies from a single cell line.
Wide range of applications: These antibodies produced through hybridoma technology have a wide range of applications in diagnostics, therapeutics, and research.
Cost-effective: Hybridoma technology is cost-effective compared to other methods used for producing monoclonal antibodies.

Process of hybridoma formation

Monoclonal antibody production

Monoclonal antibodies, also called (Mabs) are proteins made in laboratories that act like proteins called antibodies in our bodies. They seek out foreign materials and stick to them to destroy them. Laboratory-made monoclonal bodies help to stimulate our immune system.

Steps

The industrial production of monoclonal antibodies starts in the laboratory by inserting an antigen into a molecule that can trigger an immune response in a mouse.
The mouse starts to produce B-lymphocytes which are known to be antibody-producing cells.
These B-lymphocytes are produced to defend themselves from the antigen.
These cells have a short life span and fuse with immortal tumor cells (usually referred to as myeloma cells) producing so-called hybridomas. They are considered as the immortal cells that can produce antibodies.
Several B-lymphocytes clones against the antigen have been extracted from the mouse. So, hybridomas that will be able to produce various antibodies against antigens are obtained. This is a mixture of polyclonal antibodies.
Then the hybridomas coming from a single B-lymphocyte are selected, thereby obtaining an immortal cell that produces only one type of antibody. This is called monoclonal antibodies.

Monoclonal antibodies advantage

The generation of monoclonal antibodies is the desired product for clinical research. Murine antibodies are regarded as foreign when administered in humans and thus produce an immune response. This immune response against the administered Mab is called HAMA (human anti-mouse antibody response).

HAMA results in the formation of immune complexes that are rapidly cleared hence making the antibody ineffective.

Explain how hybridoma is produced

Hybridomas are produced by a process called hybridoma technology. It involves the fusion of two non-identical cells to form a hybrid cell. This technique was developed by G.Köhler and Milstein, for which they were awarded the Nobel Prize in Medicine.

The production of hybridomas requires two types of cells:

B-cells
Myeloma cells

These two cell types have unique properties that make them suitable for creating hybridomas.

These two cells are allowed to fuse and the resultant fused cell is known as a hybridoma. Each hybridoma contains genetic material from both the parent cells. It thus allows to production of antibodies like B-cells while having the ability to replicate like myeloma cells.

After fusion, the newly formed hybridomas are screened and selected based on their ability to produce specific antibodies against antigens. Once identified, these antibody-producing hybrids are cultured and used for various applications like diagnostic and therapeutic treatments.

Thus, the production of hybridomas plays a crucial role in biomedical research and contributes to the treatment of diseases.

Application in disease diagnosis and therapeutic uses

One of the main targets of antibiotic-based therapeutics has been the development of anti-cancer agents.
The first human-made antibody known as CDR-grafted is used clinically.
This antigen CAMPATH-1H is used as a potential therapeutic agent in B-cell lymphoma and many other inflammatory diseases.
This particular antigen CDw52 has led to a reduction in tumor remissions.

Challenges

Difficulty in Cell Fusion: One major challenge faced in hybridoma technology is achieving successful cell fusion between the antibody-producing B-cells and myeloma cells.

High cost: Hybridoma technology requires expensive equipment for producing monoclonal antibodies compared to newer techniques like recombinant DNA technology.

Hybridoma technology in vaccine development

Hybridoma technology allows for the production of large quantities of identical antibodies, which are essential components in many vaccines. This method is a more efficient and cost-effective way to produce vaccines. Vaccines developed by this method are effective as they have high specificity and are target-specific.

With hybridoma technology, scientists can produce monoclonal antibodies that specifically target certain antigens, making it possible to develop highly targeted vaccines.

For example- The first successful use of hybridoma technology in producing a vaccine was for Hepatitis B. This technology also contributed in creating flu vaccines, targeting fixed strains like H1N1 and H5N1.

Hybridoma technology vs alternative method of antibody production.

Characteristics	Hybridoma Technology	Recombinant DNA Technology
Definition	Hybridoma technology is a laboratory technique used to produce large quantities of monoclonal antibodies.	This process involves cutting and recombining DNA molecules, resulting in the creation of genetically modified organisms (GMOs).
Significance	Hybridoma technology also enables scientists to create custom-made monoclonal antibodies tailored for different purposes such as targeting disease-specific antigens or detecting biomarkers.	Recombinant DNA technology has also been utilized in industrial processes such as enzyme production and waste management.
Example	Developing disease-resistant crops	Development of genetically modified organisms.

Q&A

1. What is hybridoma technology?

Hybridoma technology is a laboratory technique used to produce large quantities of monoclonal antibodies.

2. What are two applications of hybridoma technology?

Two applications of Hybridoma technology involve:

Targeting disease-specific antigens
Detecting biomarkers.

3. What are the disadvantages of hybridoma technology

Disadvantages include

Contamination: There is an increased risk of contamination from bacteria or other microorganisms as cells are cultured for longer periods. It could damage the quality and specificity of produced antibodies.
High-cost effectivity
Time-consuming

Summary

Hybridoma technology is a laboratory technique used to produce large quantities of monoclonal antibodies
Monoclonal antibodies, also called (Mabs) are proteins made in laboratories that act like proteins called antibodies in our bodies.
They seek out foreign materials and stick to them to destroy them.
Laboratory-made monoclonal bodies help to stimulate our immune system.
This technology has proved capable of producing rodent antibodies of predetermined specificity to a wide variety of different antigens.
Hybridoma technology allows the production of hybrid cell lines from B cells that secrete a single, monoclonal antibody with one binding specificity.
With hybridoma technology, scientists can produce monoclonal antibodies that specifically target certain antigens, making it possible to develop highly targeted vaccines.
The first human-made antibody known as CDR-grafted is used clinically.
This antigen CAMPATH-1H is used as a potential therapeutic agent in B-cell lymphoma and many other inflammatory diseases.
This technology also contributed in creating flu vaccines, targeting fixed strains like H1N1 and H5N1.
One major challenge faced in hybridoma technology is achieving successful cell fusion between the antibody-producing B-cells and myeloma cells.
It requires expensive equipment for producing monoclonal antibodies compared to newer techniques like recombinant DNA technology.

References

Written By: Sushmita Mukhopadhyay