Research On Development Of Monoclonal Antibody Against Chicken IgM And Its Application

The chiken-breeding become a big channel of civil earning in the latest years. Disease is the biggest henace to lags-breeding, and infectious disease is the seriousest disease, account for 75 percent of all. Especialy the acute deadly infection just like Avian Influenza, since long ago it has outbroken in many countries and areas and has resulted in large economic loss, it is a severe disease devastating the stock-breeding and human health.The diagnosis of chiken infection is dependent on the isolation and characterization of the AIV, serology analysis, molecular diagnosis and electronic microscope technique. Serology tests include HA, HI, NIT, AGP, NT, ELISA and IFT. However, these methods have the disadvantages of time consuming and expensive lab equipment cost which make them not suitable for clinical instant diagnosis. Therefore, it is essential to establish a new fast diagnosis method. In this paper an instant method is successfully established to detect IgM using indirect ELISA assay and will be very important.In this paper BALB/c mice are immunized with rarefild chiken IgM oil emulsion vaccine for a long time. Then spleen cells from mouse which have high antibody titer are hybridized with SP2/0 bone marrow tumor cells with PEG-1500. The hybridization rate is 76.25％. An ELISA method is established to detect the McAb in the supernatant of the hybrid cell culture. Preliminary test shows a positive rate of 16.39％. After several sub-cloning and repetitive selection one hybrid cell lines are acquired namely 1F7 stably expressing anti-chiken IgM McAb. And it can secrete high titer antibody above 1:1000 after repetitive frozen storage, thawed revival and generation, The hybridoma cells are injected into the mice abdomen and seven days later ascites water is collected to purify the McAb by caprylic-sulfate ammonium method. The titers are above 1:32,000. Preliminary biologic characterization of the McAbs shows that the McAbs are highly affinitive and specific. There are further proved to be IgG1,κsubtype by latex agglutination test. It is estimated that they will play important roles in the diagnosis of chiken IgM.In this paper 5-10 ages chiks are immunized with H5 strain derived AIV oil emulsion antigen vaccine for one time. And then collected blood serum from third day to tenth day, detected the titers of IgM by indirect ELISA.Conclusions: In this paper anti-IgM polyclonal antibody and McAb are successfully prepared and used in the diagnosis of determination of the state of bacterin immunity and the detection of IgM. It is shown that indirect ELISA using anti-IgM McAb has the advantages of high sensitivity and specificity, good repeatability and handling and on-line monitoring of the detecting process.So anti-IgM McAb prepared in this research offer a current reagent for research of chicken infection.