Question:
Which of the following combination is appropriate for hybridoma production?
Which of the following combination is appropriate for hybridoma production?
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Hybridoma = Splenocytes (immune mouse B-cells) + Myeloma (immortal tumor cells) selected on HAT medium. PEG is simply the chemical glue (fusogen) used to fuse them, not a cell partner or selective agent.
Updated On: Aug 7, 2026
- Lymphocytes from immune mice, HAT, Myeloma cells
- Lymphocytes from immune mice, HAT, PEG
- Sendai virus, HAT, Myeloma cells
- Lymphocytes from immune mice, Aminopterin, Myeloma cells
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The Correct Option is A
Solution and Explanation
Step 1: Understanding the Concept:
Hybridoma technology, developed by Georges Köhler and César Milstein in 1975, is used to produce monoclonal antibodies on a large scale.
It involves fusing short-lived antibody-producing cells with immortal cancer cells and selecting for successfully fused hybrid cells.
Step 2: Detailed Explanation:
The generation of a functional hybridoma requires three essential components:
1. Antibody-producing B-lymphocytes: These are isolated from the spleen of a mouse immunized with the target antigen.
These cells produce the desired antibody but have a limited lifespan in culture.
2. Myeloma cells: These are immortal, cancerous B-cell lines that lack the enzyme hypoxanthine-guanine phosphoribosyltransferase ($\text{HGPRT}^-$), making them unable to use the salvage pathway for DNA synthesis.
3. HAT Selection Medium: After cell fusion (facilitated by polyethylene glycol or PEG), the mixture contains fused hybrids, unfused lymphocytes, and unfused myeloma cells.
- Unfused lymphocytes die naturally due to their short lifespan.
- Unfused myeloma cells are killed by the aminopterin in HAT medium because they lack HGPRT and cannot synthesize DNA.
- Only the hybridoma cells survive because they inherit immortality from the myeloma parent and the functional HGPRT gene (salvage pathway) from the lymphocyte parent.
Therefore, the appropriate combination is lymphocytes from immune mice, HAT medium, and myeloma cells.
Step 3: Final Answer:
The essential combination for hybridoma production corresponds to option (A).
Hybridoma technology, developed by Georges Köhler and César Milstein in 1975, is used to produce monoclonal antibodies on a large scale.
It involves fusing short-lived antibody-producing cells with immortal cancer cells and selecting for successfully fused hybrid cells.
Step 2: Detailed Explanation:
The generation of a functional hybridoma requires three essential components:
1. Antibody-producing B-lymphocytes: These are isolated from the spleen of a mouse immunized with the target antigen.
These cells produce the desired antibody but have a limited lifespan in culture.
2. Myeloma cells: These are immortal, cancerous B-cell lines that lack the enzyme hypoxanthine-guanine phosphoribosyltransferase ($\text{HGPRT}^-$), making them unable to use the salvage pathway for DNA synthesis.
3. HAT Selection Medium: After cell fusion (facilitated by polyethylene glycol or PEG), the mixture contains fused hybrids, unfused lymphocytes, and unfused myeloma cells.
- Unfused lymphocytes die naturally due to their short lifespan.
- Unfused myeloma cells are killed by the aminopterin in HAT medium because they lack HGPRT and cannot synthesize DNA.
- Only the hybridoma cells survive because they inherit immortality from the myeloma parent and the functional HGPRT gene (salvage pathway) from the lymphocyte parent.
Therefore, the appropriate combination is lymphocytes from immune mice, HAT medium, and myeloma cells.
Step 3: Final Answer:
The essential combination for hybridoma production corresponds to option (A).
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