[Generation of human monoclonal antibodies against hemorrhagic fever with renal sydrome (HFRS) virus].
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.
Zhao
H L
HL
Military Medical Institute of Shenyang Military Region, Shenyang 110031.
Zang
J
J
Li
Z Y
ZY
Zhang
X
X
Jiang
S
S
Liu
J
J
Wei
X
X
He
Y
Y
D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D018063:Hantaan virus / Q000276:immunology*; D006480:Hemorrhagic Fever with Renal Syndrome / Q000276:immunology* / Q000821:virology; D006801:Humans; D006825:Hybridomas / Q000276:immunology
D000911:Antibodies, Monoclonal