Hybridoma production by simplified avidin-mediated electrofusion.
A simplified method for producing cell hybrids by avidin-mediated electrofusion has been developed. First, biotin was attached both to immunogen and to the surface of murine myeloma cells using N-hydroxysuccinimidobiotin. Biotinylated immunogen was incubated with splenocytes derived from immunized mice and allowed to bind to surface immunoglobulins of B cells. Using streptavidin, biotinylated myeloma cells then were bridged to those B cells bearing biotinylated immunogen. Selective fusion of bridged cells was accomplished by their limited exposure to high-voltage potentials. A high frequency of hybridomas was obtained all of which secreted high titers of antibodies to a selected model immunogen, keyhole limpet hemocyanin.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001360:Avidin; D001710:Biotin; D002459:Cell Fusion; D004560:Electricity; D005260:Female; D006433:Hemocyanins / Q000276:immunology; D006825:Hybridomas / Q000166:cytology*; D051379:Mice; D008807:Mice, Inbred BALB C; D010047:Ovalbumin / Q000031:analogs & derivatives*; D010954:Plasmacytoma / Q000276:immunology
D000911:Antibodies, Monoclonal; D001360:Avidin; D001710:Biotin; D010047:Ovalbumin; D006433:Hemocyanins