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Monoclonal Antibody Production (1999) / Chapter Skim
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Generation of Hybridomas: Permanent Cell Lines Secreting Monoclonal Antibodies
Pages 8-11

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From page 8...
... It has also been possible to genetically replace much of the mouse mAbproducing genes with human sequences, reducing the immunogenicity of mAb destined for clinical use in humans. Before the advent of the hybridoma method, investigators could produce only polyclonal serum antibodies; this required large numbers of immunized animals and did not immortalize the antibody-producing cells, so it required repeated animal use to obtain more antibodies.
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From page 9...
... Flowchart illustrating steps needed to produce mAb by mouse ascites method. Note that all steps up to production of ascites fluid are required for either in vivo or in vitro production of mAb.
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From page 10...
... Fusion is accomplished by co-centrifuging freshly harvested spleen cells and myeloma cells in polyethylene glycol, a substance that causes cell membranes to fuse. As noted in step 3, only fused cells will grow in ~ The selection growth medium contains the inhibitor aminopterin, which blocks synthetic pathways by which nucleotides are made.
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From page 11...
... Also, it is the experience of many that a brief period of growth by the mouse ascites method produces cell lines that at later in vitro and in vivo stages show enhanced hardiness and optimal antibody production (Ishaque and Al-Rubeai 1998~. Guidelines have been published to assist investigators in using the mouse ascites methods in these ways (Jackson and Fox 1995~.
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Key Terms

  • mouse ascites
  • spleen cells
  • antibody titer
  • cell fusion
  • ascites method

  • feeder cells
  • cell lines
  • growth factors
  • cell line
  • hybridoma cell

  • hybridoma cells
  • injecting antigen
  • expansion method
  • product due
  • considerable judgment

  • procedure yields
  • medium contains
  • growth medium
  • vitro growth
  • culture followed

  • immunized animals
  • selection medium
  • inhibitor aminopterin
  • various techniques
  • repeated animal

  • bypass pathway
  • prepared myeloma
  • immortalized cells
  • versus total
  • commercial preparations


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