Hybridoma Technology for the Production of Monoclonal Antibodies

Summary

This video explains the process of creating monoclonal antibodies using hybridoma technology. It begins by introducing monoclonal antibodies, which are derived from a single B-cell and bind to a specific epitope, unlike polyclonal antibodies. The hybridoma technique involves injecting an antigen into a mouse, isolating B-cells, and fusing them with immortal myeloma cells to produce hybridomas. These hybridomas can be cultured and cloned to generate monoclonal antibodies, which can be further tested for specificity using techniques like ELISA and Western blot. The video also encourages viewers to subscribe and supports the channel.

Takeaways

• 😀 Hybridoma technology is used to produce monoclonal antibodies, which are identical and bind to a single specific epitope on the antigen.
• 😀 Monoclonal antibodies differ from polyclonal antibodies, which bind to multiple epitopes from various B cells.
• 😀 Monoclonal antibodies are often used therapeutically, such as in the treatment of certain types of cancer.
• 😀 In the first step of hybridoma technology, an antigen is injected into a mouse, triggering an immune response.
• 😀 B cells in the mouse recognize the foreign antigen and begin producing antibodies against it.
• 😀 Plasma cells, which are found in the spleen, are isolated and cultivated in cell culture to produce antibodies.
• 😀 A major challenge is that plasma cells are not immortal, meaning they cannot be cultured indefinitely.
• 😀 To overcome this, plasma cells are fused with immortal myeloma cells, creating hybridoma cells.
• 😀 Hybridoma cells combine the ability to produce antibodies and immortality, allowing for long-term cultivation.
• 😀 These hybridoma cells are cultured in a selective medium that only supports growth of cells with both characteristics.
• 😀 Single hybridoma cells are isolated and grown into clones, producing monoclonal antibodies, which can be further tested using techniques like ELISA or Western blot.

Q & A

What are monoclonal antibodies and how do they differ from polyclonal antibodies?

Monoclonal antibodies are identical antibodies produced from a single B cell, and they bind to a single specific epitope on an antigen. In contrast, polyclonal antibodies are produced by multiple B cells of different origins and can bind to various epitopes on the same antigen.

Why are monoclonal antibodies often used in therapeutics?

Monoclonal antibodies are used in therapeutics because they are highly specific, targeting only one particular epitope on an antigen. This makes them effective in treatments such as cancer therapies, where precision is crucial.

What is the first step in generating monoclonal antibodies using hybridoma technology?

The first step involves injecting the antigen of interest into a mouse, triggering an immune response. This leads to the production of antibodies by the mouse's B cells, which recognize and respond to the foreign protein.

What are splenocytes, and why are they important in the hybridoma process?

Splenocytes are plasma cells found in the spleen of the mouse. They are essential in hybridoma technology because they produce antibodies in response to the antigen, but they cannot grow indefinitely, which is why they need to be immortalized.

What role do myeloma cells play in hybridoma technology?

Myeloma cells are immortal cancer cells that can grow indefinitely. When fused with plasma cells from the mouse, they create hybridoma cells that can produce specific antibodies and grow indefinitely.

How are hybridoma cells created?

Hybridoma cells are created by fusing plasma cells (splenocytes) from the mouse with myeloma cells. This fusion creates cells that can both produce specific antibodies and grow indefinitely.

Why is a selective medium used in culturing hybridoma cells?

A selective medium is used to ensure that only hybridoma cells, which have the ability to produce antibodies and grow indefinitely, survive and proliferate. It eliminates cells that do not have both characteristics.

What is the significance of isolating single hybridoma cells?

Isolating single hybridoma cells is important to ensure that each well contains clones of cells derived from a single hybridoma. This way, the antibodies produced are monoclonal and originate from one single cell.

How can the specificity of monoclonal antibodies be tested?

The specificity of monoclonal antibodies can be tested using techniques like ELISA (Enzyme-Linked Immunosorbent Assay) or Western blotting. These methods help confirm that the antibodies bind only to the intended antigen.

What is the importance of monoclonal antibodies in cancer treatment?

Monoclonal antibodies are important in cancer treatment because they can be designed to target specific antigens present on cancer cells, allowing for more precise treatment with fewer side effects compared to traditional therapies.