Stable Antibody-Producing Murine Hybridomas

Abstract

A method is described for obtaining antibody-producing hybridomas that are preferentially retained in cultures of fused mouse spleen and myeloma cells. Hybridomas are produced by fusing mouse myeloma cells that are deficient in adenosine phosphoribosyltransferase (APRT) with mouse spleen cells containing Robertsonian 8.12 translocation chromosomes. The cell fusion mixtures are exposed to a culture medium that can be utilized only by APRT-positive cells, which results in the elimination of both unfused APRT-deficient myeloma cells and non-antibody-producing APRT-deficient hybridomas that arise by segregation of the 8.12 translocation chromosomes containing the APRT genes and the active heavy chain immunoglobulin gene.