Calibration of hybridoma antibody assays by polyclonal supernatants produced in vitro.
Conventional antisera used as a positive control in antibody tests during hybridoma screening and processing can be replaced to advantage by supernatants of short term spleen cell cultures. Such cultures can easily be initiated from the same spleen cell suspension which is prepared in connection with a fusion experiment. Either supernatants of corresponding cultures from nonimmune animals or medium alone can serve as a negative control. In vitro produced polyclonal standards show very low background binding compared to antisera raised in vivo. They are convenient "ready-to-use" reagents with an antibody titer comparable to that expected in hybridoma culture fluids. There is no need for immunization and bleeding of separate animals. In addition, these supernatants provide an internal control of the immunization efficiency.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D002478:Cells, Cultured; D005455:Fluorescent Antibody Technique; D006825:Hybridomas / Q000276:immunology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D012016:Reference Values; D013154:Spleen / Q000276:immunology; D013997:Time Factors
D000911:Antibodies, Monoclonal