Peripheral blood lymphocytes (PBL) were obtained from healthy donors and treated with L-leucy-leucin methy (LeuLeu-OMe). The LeuLeu-OMe treated PBL were in vitro immunized with HFRS virus proteins (55kD, 67kD) for 6 days, and then fused with K6H6/B5 cells in medium containing 45% (PEG) and 7.5% DMSO. The fused cells were seeded in 96-well plates at cell density 0.5-1 x 10(5) cells/well in HAT supplemented RPMI-1640 medium. After 14-20 days, 3 positive wells were deteced by ELISA and two (1B7, 2D5) of the 3 hybridoma were stable after 4 time cloning. The hybridoma 2D5 appears to be with high ability of production of the antibodies at a concentration of 20-30 microg/ml/10(6) cells/24h. It was proved by IFAT that 2D5 McAb was specific for HFRSV.
[Generation of human monoclonal antibodies against hemorrhagic fever with renal sydrome (HFRS) virus] Publishing Authors By Initials
