Abstract
A fusion protocol was designed for the optimal production of hybridomas following electrofusion of human B cells with cells of the heteromyeloma fusion partner SPAM-8. Peripheral blood lymphocytes showed an average fusion efficiency of 0.4 x 10-4 whereas Epstein-Barr virus-transformed B cells showed fusion efficiencies ranging from 6.2 x 10-4 to 9.0 x 10-4. Similar results were obtained with bone marrow-derived lymphocytes. Trypsin treatment of the cells prior to electrofusion further increased the fusion efficiency to 12.3 x 104. In comparison, conventional polyethylene glycol-induced fusion resulted in a fusion efficiency of 0.8 x 10-4. Thus, electrofusion of human B cells with SPAM-8 heteromyeloma cells introduced a 15-fold increase in hybridoma formation as compared to the conventional fusion method.
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