[Screening and identification of monoclonal antibodies against human neutrophil and detection of inhibiting opsonic phagocytic activity].
Using Ficoll-Hypaque gradient centrifugation, we have successfully obtained the neutrophils from normal human donors, which can be used as particle antigens for immunizing BALB/c mice. Hybridomas were produced through the fusion of SP2/0 myeloma cells and splenocytes from immunized BALB/c mice. The ratio of fusion was 1:5. The cells were fused with 30% polyethylene glycol (MW 4000). The rate of fusion was 90%. The antibody producing colonies against the membrane of neutrophils in HAT medium were selected by indirect immunofluorescence assay. Antibody positive rate was 60% (102/170). Limiting dilution was used for colonization of the antibody-producing hybridoma cells. After colonization for three times, one hybridoma cell line was obtained, which could inhibit complement-mediated phagocytosis. Culture supernatant was used against class- and subclass- specific rabbit antisera (rabbit anti-mouse IgM, IgG, IgG1, IgGd22, IgG2b, IgG3) in an agar immunodiffusion system, it was shown to be IgG22.
Xu
L
L
Gao
J
J
Wang
B
B
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D006801:Humans; D006825:Hybridomas; D007074:Immunoglobulin G / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D009504:Neutrophils / Q000276:immunology*; D009895:Opsonin Proteins / Q000276:immunology; D010587:Phagocytosis / Q000276:immunology*
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D009895:Opsonin Proteins