Bioinformatics for Vet. Part XXV. Sung Youn Lee, PhD. Student Veterinary collage, Room 320 02 450 3719, 016 293 6059 [email protected]. MCA: Monoclonal Ab. MIX and Fusion Screen Production. Step 1. Mix spleen cells from a mouse that has been immunized with the desired antigen with - PowerPoint PPT Presentation
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Bioinformatics for Vet. Part XXVBioinformatics for Vet. Part XXV
• Mix – spleen cells from a mouse that has been immunize
d with the desired antigen with – myeloma cells.
• Use an agent to facilitate fusion of adjacent plasma membranes. (PEG-polyethylenglycol or Sendai virus). Even so, the success rate is so low that there must be a way to select for the rare successful fusions.
Step 2.
• So use myeloma cells that have: – lost the ability to synthesize hypoxanthine-guanine-
phosphoribosyltransferase (HGPRT). This enzyme enables cells to synthesize purines using an extracellular source of hypoxanthine as a precursor. Ordinarily, the absence of HGPRT is not a problem for the cell because cells have an alternate pathway that they can use to synthesize purines.
– However, when cells are exposed to aminopterin (a folic acid analog), they are unable to use this other pathway and are now fully dependent on HGPRT for survival.
– lost the ability to synthesize any antibody molecules of their own (so as not to produce a hybridoma producing two kinds of antibody molecules).
8F2-This monoclonal reacts with antigenic group II viruses of the family Coronaviridae, thus reacting with bovine coronavirus and elk coronavirus nucleoproteins. However, this monoclonal does not recognize antigenic group I virus such as transmissible gastroenteritis virus (TGEV).
Thank you for your attention ~Thank you for your attention ~