[Studies on monoclonal antibody against recombinant human granulocyte colony-stimulating factor]

Abstract

Three hybridomas producing monoclonal antibodies (McAb) against recombinant human granulocyte colony-stimulating factor (rhG-CSF) have been established by fusing mouse myeloma SP 2/0 cells with spleen cells from a BALB/c mouse immunized with rhG-CSF. Ascites was produced from BALB/c mice by injecting substrain 2D4-B4 hybridoma cells intraperitoneally. The antibody was purified either by the caprylic acid-ammonium sulfate method or by euglobulin precipitation. The caprylic acid-ammonium sulfate method was superior to euglobulin precipitation in terms of purification and recovery of IgG. The 2D4-B4 McAb was determined to belong to the IgG3 subclass with a molecular weight of about 172,400 It was proved by Western Blot to react specifically with rhG-CSF. The stability and affinity of the McAb were analyzed as well. The potential clinical and experimental applications of this McAb are discussed.