In Vitro Expression Of IgMμ Chain Of Chicken, Porcine And Bovine And Preparation Of Monoclonal Antibody Against IgMμ Chain

With the development of intensive animal production, infectious diseases of domestic animals and fowl have jeopardized the intensive animal industry seriously. Infectious diseases not only cause a great number of animals to die and a considerable production loss, but also are harmful to people health. In order to control the condition of illnesses, condemn the infected animals and degrade the production losses, it is urgent to set up an early and quick method to diagnostic infectious disease.IgM is the premature immunoglobulin in the first humoral immune reaction and plays a significant part in anti-infection immunity. It is importmant to do early diagnosis by means of IgM detection. BALB/c mice were immunized four times with prokaryotic expressed protein of IgM μ chain. The spleen cells of the hyperimmunized mice and SP2/0 cells were fused with PEG4000. The cell fusion rates were all above 90%. An indirect ELISA coated with protein expressed in recombinant Baculovirus was developed to detect monoclonal antibodies(mAbs) secreted by hybridoma cell lines. After three successive subclones, one hybridoma cell strain, named by 1H2, secreting specific mAbs for chicken IgM μ chain was obtained using hybridoma techniques. After frozen storage, thawed revival and generation repeatedly, the hybridoma cell strain still produced high titer mAbs steadily. By means of immunoglobulin subclass analysis, the obtained mAbs was classified as IgG₁ κ . The ascities fluids titers of the mAb was 1:1 × 10⁶. The results of indirect ELISA and Western-blot showed that the mAbs can be used to detect IgM in chicken, porcine and bovine serium quickly, sensitively and specially.It is believed that the specific mAbs for chicken IgM μ chain will supply a new convenient method for early diagnosis of infectious diseases and be useful to prepare anti-IgM μ, monoclonal antibody kits. It can also be used to detect the cells containing IgM in tissues and peripheral blood quantitatively. It can be used to study the angtigen presentating...