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 micrograms/ml of culture medium, whereas IgM production of EBV-transformed B cell clones ranged between 3.0 and 4.0 micrograms/ml.
|Autori interni:||GARZELLI, CARLO|
|Autori:||GARZELLI C; PUGLISI C; FALCONE G|
|Titolo:||HUMAN MONOCLONAL-ANTIBODY TO PURIFIED PROTEIN DERIVATIVE OF TUBERCULIN PRODUCED BY HYBRIDS CONSTRUCTED WITH EPSTEIN-BARR VIRUS-TRANSFORMED LYMPHOCYTES-B AND MOUSE MYELOMA CELLS|
|Anno del prodotto:||1986|
|Digital Object Identifier (DOI):||10.1002/eji.1830160522|
|Appare nelle tipologie:||1.1 Articolo in rivista|