Outline the key steps involved in producing a monoclonal antibody against human interleukin‑2 (IL‑2) using hybridoma technology. List the procedures from immunization to screening.
Answer
Immunize mouse → fuse spleen cells with myeloma cells → select fused cells (HAT medium) → screen supernatants for IL‑2 binding → clone positive hybrids → expand and purify antibodies.
Step-by-step solution
- Immunize a BALB/c mouse with purified human IL‑2 protein to elicit specific B‑cell response.
- Harvest spleen cells from the mouse and fuse them with an immortalised myeloma cell line (e.g., Sp2/0) using polyethylene glycol.
- Plate fused cells in HAT medium to select for hybridoma cells that have both antibody production capability and immortality.
- Screen hybridoma supernatants by ELISA to identify clones that bind specifically to IL‑2.
- Clone positive hybrids by limiting dilution to obtain monoclonal population.
- Expand selected clones in culture and purify the secreted monoclonal antibody using protein G chromatography.
Common mistake: Forgetting the HAT selection step
Concept tested: Hybridoma monoclonal antibody production
Published · How this solution was prepared and checked
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