Preparation Monoclonal Antibody Of β-mannanase And Studies On Methods Of The Detecting β-mannanase

The β-mannanase from trichoderma reesei RutC-30 is induced by gum locust bean and lactose through 10L fluid ferment, and is purified by twice (NH4)2SO4 precipitation and twice PAGE purification, getting purified ?-mannanase. The molecular weight of the enzyme is 53KD. Female Balb/c mice were immunized by purified β-mannanase, the spleen cell of immunized mice was fused with myeloma cell SP2/0 by using 50% PEG. A hybridoma which secreted anti ?-mannanase Mcab with high specialization was selected by ELISA screening positive hybridomas using HAT. The Monoclonal Antibody from the mouse ascites was first purified by (NH4)2SO4 precipitation. The ?-mannanase from trichoderma reesei is used as antigen to immunize male rabbit produce anti- ?-Mannanase polyclonal antibodies and e stablish the ELISA double s andwich method for detecting product expressed transform ?-mannanase gene E.coli. To establish method to detect product expressed in E.coli of pig bowel.