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Microbiology · Immunology: hybridoma selection
Sample itemIn producing monoclonal antibodies, mouse spleen B cells are fused with HGPRT-deficient myeloma cells and cultured in HAT medium. Why do only the hybridoma cells survive?
Original practice item, not a recalled exam question. Nothing here is recorded.
The answer and the reasoning
D. Myeloma cells cannot use the salvage pathway when aminopterin blocks de novo synthesis
Aminopterin blocks the de novo purine and pyrimidine pathway, so cells must use the salvage pathway with hypoxanthine and thymidine, which needs HGPRT. Unfused myeloma cells lack HGPRT and die, and unfused spleen cells die because they have a limited life span in culture. Hybrids inherit HGPRT from the spleen cell and immortality from the myeloma cell.
Option by option
A. Hypoxanthine is toxic to unfused myeloma cells
Hypoxanthine is the substrate of the salvage pathway, not a toxin.
B. Thymidine allows spleen B cells to grow indefinitely in culture
Thymidine is a salvage pathway substrate and does not confer immortality.
C. Aminopterin selectively kills spleen B cells but spares myeloma cells
Aminopterin acts on all cells through the de novo pathway and does not discriminate spleen cells from myeloma cells.
D. Myeloma cells cannot use the salvage pathway when aminopterin blocks de novo synthesis (correct)
The enzyme defect leaves myeloma cells unable to salvage nucleotides once the de novo route is blocked.
Source note: Original NextPrep sample item — pending expert review. This item is original practice material awaiting independent expert review, so confirm important facts in a standard textbook.
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