Outline the process by which hybridoma cells are made
What will be an ideal response
Mice are immunized with the desired antigen. Their spleens are harvested and homogenized to release B cells. The
B cells are mixed with multiple myeloma cells in the presence of polyethylene glycol which promotes fusion of the
cell membranes. The resultant hybridoma cells are selected by using a differential culture medium. The myeloma
cells must be nonantibody producing and HGPRT negative (unable to convert hypoxanthine to guanine). Normal B
cells are HGPRT positive. If the hybridoma cells are grown in medium that inhibits synthesis of nucleotides and
forces cells to use the hypoxanthine pathway, then only B cells and successfully fused hybridoma cells can grow.
The B cells eventually die out, leaving only hybridoma cells producing the antibodies of the B cells fused in them.
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