A new approach to the generation of human or murine antibody producing hybridomas.

A new and very efficient method for the generation of human and murine monoclonal antibodies has been developed. The method is based on clonal expansion of single B cells in the presence of human T cell supernatant and irradiated murine thymoma helper cells. Subsequently, the clonally expanded B cells are immortalized by electric field mediated cell fusion. The high efficiency of the method permits the processing of small numbers of lymphocytes, e.g., obtained by preselection of specific B cells, small amounts of human donor material and murine PBL or lymph node cells. The method may be an alternative for the EBV-transformation technique used for the generation of human monoclonal antibodies, which immortalizes only a subset of B cells and frequently yields poorly growing or unstable cell lines. This report describes the generation of murine anti-HIV and human anti-rubella antibodies combining the clonal expansion of B cells and mini-electrofusion.