Human monoclonal antibody to purified protein derivative of tuberculin produced by hybrids constructed with Epstein‐Barr virus‐transformed B lymphocytes and mouse myeloma cells

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Abstract
A method of producing human monoclonal antibody by combining somatic cell hybridization technology with the capability of Epstein‐Barr virus (EBV) to transform human B lymphocytes is described.Peripheral blood lymphocytes from a donor with positive tuberculin skin test reaction were transformed by EBV and then tested for antibody production to mycobacterial purified protein derivative (PPD) by an enzyme‐linked immunosorbent assay. Two EBV‐transformed lymphoblastoid cell lines making IgM antibodies to PPD were obtained. One of these cell lines was fused by polyethylene glycol with a murine hypoxanthine‐guanine phosphoribosyl transferase‐deficient myeloma cell line that had been selected for resistance to ouabain. The human‐mouse hybrids were selected in ouabain‐containing HAT medium and 11 heterohybridomas producing IgM antibody to PPD were obtained. one of these was cloned by limiting dilution with efficiency at least 20‐fold higher than parent EBV‐transformed cell line. Heterohybridoma subclones reached levels of IgM antibody as high as 75.0 μg/ml of culture medium, whereas IgM production of EBV‐transformed B cell clones ranged between 3.0 and 4.0 μg/ml.