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.