An山gen Myeloma cells (HGPRT-.Ig-,"immortal") Primed B-cells (HGPRT+,le+,“ota) Polyethylene glycol Unfused B cell Unfused myeloma cell Heterokaryons Dies in HAT HAT selection (only B-cell/myeloma hybridoma grows) Hybridoma (containing genes for HGPRT+,Ig+,immortal growth) Assay for desired antibody in culture supernatant Reclone Ab+ hybridomas Expand positive clones Culture Ascites fluid Monoclonal antibodies Monoclonal antibodies