THE ESTABLISHMENT OF HYBRIDOMA-CELL-LINES PRODUCING MONOCLONAL ANTIBODIES AGAINST UROKINASE / 第二军医大学学报

ABSTRACT

Spleen cells of BALB/C mice immunized with medical urokinase were fused with FO and NS-1 myeloma cells three times, and six hybridoma cell lines(I-UB5, I-UD2, 3-UA2, 3-UD12,3-UG2 and 3-UH7)secreting monoclonal antibodies against urokinase were obtained after screening.Of them, I-UB5 and I-UD2 v/ere cloned, frozen, stored, resuscitated and analysed chromosomally.It was found that I-UB5 and I-UD2 were clones obtained by fussing spleen cells of BALB/C mice with FO myeloma cells.Their subclasses are IgG1 and IgG2a respectively determined by immunodifussion and enzyme-linked immunosorbent assay(ELISA).The titers dctc-termined by ELISA were in the range of 10-2-10-3 in culture supernatants, and 10-6-10-7 in ascitic fluid.The I-UB5 and I-UD2 were proved to be monoclonal antibodies against high molecular weight subunit of urokinase, since they did not react with the purified low molecular weight subunit of, but reacted with mixed subunit of urokinase