Clostridium Chauvoei Antigen Purification And Analysis

This experiment was conducted to understand the characteristics of emphysematous gangrene Clostridium antigen extract after anaerobic culture of emphysematous gangrene Clostridium bacteria and secretion of toxins, the crude antigen for emphysema gangrene Clostridium standard strains (C54-1) after ultrasonic lysis prepared from the crude secretory antigen prepared from processing by the toxin by saturated ammonium sulfate, and then through Sephadex G-200gel column separation and purification, the application of vertical gel electrophoresis (SDS-PAGE). and immunoblotting experiments(Western blotting) emphysematous gangrene Clostridium crude reference to the purified somatic antigen and secrete antigen reactogenicity analysis. The results showed that emphysematous gangrene Clostridium antigen eight apparent protein peak, polyacrylamide gel electrophoresis, respectively, corresponding to the bands, the molecular mass of93kD,72kD,56kD,46kD,38kD,26kD,15kD,11kD, of which the second (72kD), five (38kD), the peak in six (26kD) antigen showed significant specificity. Secretory antigen of seven major protein peak, the application of vertical gel electrophoresis (SDS-PAGE)., Showing the blue stripe is the second, third, fifth, seventh protein peak, protein molecular weight, respectively, for the93kD70kD,48kD and32kD, immunoblotting analysis of emphysematous gangrene component of bacterial antigens and secreted protein antigens found positive serum of mice infected with emphysema, gangrene Clostridium reacts with the antigen component proteins, including bacterial antigen72kD,38kD,26kD protein band color deepest secretory antigen93kD and48kD showing a specific band.Purification of specific bacteria and secretion of antigens by adding the adjuvant, the immunogenicity of the immune mice (25g/only), and the addition of the control group, using the agar diffusion assay principle, emphysematous gangrene Clostridium-specific protein antigens purified do Analysis. The results showed that the highest antibody titers produced by somatic antigen of the specific emphysematous gangrene Clostridium1:64, the whole-cell antigen titer crude extract; emphysematous gangrene Clostridium-specific secretory antigen produced by The antibody titer can reach up to1:64, than the pure bacterial culture supernatant produced by high titer. This experiment illustrates the purification of specific bacteria and secretion of antigen with good immunogenicity. This study identified the reactogenicity and immunogenicity of the antigen in the bacterial antigen and secrete antigen in emphysematous gangrene Clostridium, to provide experimental basis for the later emphysematous gangrene serological diagnostic methods to establish and vaccine development.