Hybridoma technology for monoclonal antibodies: which components are essential to generate stable, antibody-secreting clones?
-
AMouse splenic lymphocytes (antibody-producing B cells)
-
BMouse myeloma cells (immortal fusion partners)
-
CBoth mouse splenic lymphocytes and mouse myeloma cells
-
DNone of these
-
EOnly human peripheral blood mononuclear cells
Answer
Correct Answer: Both mouse splenic lymphocytes and mouse myeloma cells
Explanation
Introduction / Context:Monoclonal antibody (mAb) production via hybridoma technology fuses a specific antibody-producing B cell with an immortal myeloma cell, yielding a hybrid that both secretes a single antibody and proliferates indefinitely. Recognizing the required inputs is foundational in immunotechnology.
Given Data / Assumptions:
- Splenic B cells provide antibody specificity.
- Myeloma cells provide immortality and selective growth traits (e.g., HGPRT deficiency for HAT selection).
Concept / Approach:After immunizing a mouse with an antigen, spleen cells are harvested and fused with compatible myeloma cells using PEG. Hybrids are selected in HAT medium; positive clones are screened for desired specificity and subcloned to monoclonality.
Step-by-Step Solution:
1) Identify the source of specificity: antigen-primed splenic B cells.2) Identify the source of immortality: myeloma fusion partners.3) Conclude both cell types are necessary to create stable monoclonal-producing hybridomas.Verification / Alternative check:Köhler and Milstein’s seminal approach underpins modern monoclonal antibody production and biotech manufacturing.
Why Other Options Are Wrong:
- Either cell alone is insufficient: B cells die; myeloma cells lack specific antibody.
- “None” or “only human PBMCs”: Do not describe classical mouse hybridoma method.
Common Pitfalls:
- Confusing polyclonal sera (from whole animals) with monoclonal hybridoma products.
Final Answer:Both mouse splenic lymphocytes and mouse myeloma cells