Affiliations: Brunswick Biotechnetics, San Diego, California, USA
Note: [] Address requests for reprints to: Martin I. Mally, Brunswick Biotechnetics, 4116 Sorrento Valley Boulevard, San Diego, CA 92121, USA.
Abstract: The production of human monoclonal antibodies is limited by a lack of good human fusion partners, the short-term stability of most human hybridomas, low fusion frequencies, and low secretion levels. An alternative method of immortalizing human immunoglobulin-secreting lymphocytes is presented here, which avoids many of the drawbacks associated with human hybridoma technology. Brief exposure of cells to a high intensity electric field results in the reversible permeabilization of cell membranes (electroporation). This technique allows the introduction of genes into virtually any cell type in a highly efficient manner. Specific genes or DNA sequences can thus be transferred into cells via the transient pores created in the membrane and, if the sequences are expressed, can yield cells with altered phenotypic properties (i.e., drug resistance). Electroporation of activated human lymphocytes in the presence of DNA sequences coding for immortalization functions and specifically targeted to B cells, would yield, via the stable integration of exogenous DNA into the cellular genome and the subsequent expression of the genes, immortalized, immunoglobulin-secreting lymphocytes, without the need for a fusion partner. Details of this technique, in addition to its advantages over the commonly used methods for generating hybridomas, are discussed.
