Study On Targeting Actinobacillus Serum Antibody And Amikacin Complexes Targeting Study

In order to conjugate Porcine Contagious Pleuropneumonia actinomycetes serum antibody and Amikacin sulfate,and Coupling will not make pharmacodynamics of Amikacin sulfate and biological activity of Porcine Contagious Pleuropneumonia actinomycetes serum antibody Lost or severely reduced. This experiment first expected to the isolation and identification of Porcine Contagious Pleuropneumonia in actinomycetes from the disease and maked oil emulsion adjuvant inactivated vaccine. Then used the preparation of oil emulsion adjuvant inactivated vaccine on many occasions by the shallow intramuscular immunization of healthy rabbits.Then separated serum after21d. Detected antibodies against Porcine Contagious Pleuropneumonia actinomycetes serum antibody titer by indirect ELISA. Serum antibody titer of indirect ELISA is1:2048. Then screened Porcine Contagious Pleuropneumonia actinomycetes serum antibody and Amikacin sulfate coupling condition. With different concentrations of Amikacin sulfate. serum antibody. PEG6000. build square test. Through the comparison of screening out coupling yield the best formula is Amikacin sulfate600(mg):serum antibody in3(g):PEG600015(g). Observation results of Gram staining, polyacrylamide gel electrophoresis, and electron microscopic of conjugates displayed good conjugates of Amikacin sulfate and serum antibody.In order to explore the conjugates for targeted, this experiment detected conjugates with indirect ELISA、indirect immunofluorescence and immunoelectron microscopic. Indirect ELISA test shows that Coupling of antibodies without influence of antigenicity. Immunofluorescence and immunogold electron microscopy revealed that Conjugates have specific targeting against Porcine Contagious Pleuropneumonia actinomycetes. Conjugates and Porcine Contagious Pleuropneumonia actinomycete bacteria liquid mixed3min. conjugates and Porcine Contagious Pleuropneumonia actinomycetes combinated:Conjugates and Porcine Contagious Pleuropneumonia actinomycete bacteria liquid mixed7min, Conjugates could enter the Porcine Contagious Pleuropneumonia actinomycete bacteria in vivo.However, serum antibody and Porcine Contagious Pleuropneumonia actinomycete bacteria liquid mixed4min, serum antibody and Porcine Contagious Pleuropneumonia actinomycete bacteria combinated; Serum antibody and Porcine Contagious Pleuropneumonia actinomycete bacteria liquid mixed9min, serum antibody could enter the Porcine Contagious Pleuropneumonia actinomycete bacteria in vivo. The results of these experiments Indicated that the coupling technology can enhance the antibody targeting effect.