Preparation and immunology of conjugates of CP8 from Staphylococcus aureus

On the basis of this chemical method,we chose ADH as bridge,EDC as the couplant to bide purified CP and purified ClfA-FnBPB together in order to prepare conjugates.In the couping process to determine the best mass ratio and coupling ratio of conjugate were determind.Then the Balb/C mice were immunized with the conjugates,ELISA method and mouse toxicity test were chosen to detect antibody levels and protective effect of CP conjugates.The result showed that the conjugates were made in the CP /ClfA-FnBPB mass ratio of one to two through chemical method and the coupling ratio was 1 to 1.89.Using ELISA method,7 days after the second immunity,conjugate group showed immunogenicity,and antibody titers can up to 1∶6 400.The antibody levels did not appear in negative control group and CP group.It can stimulate immune response in fusion protein group after the second immunization,but the effect was not obvious,andantibody titers is only 1∶100.The results of mouse toxicity test showed that the conjugate group had the highest survival rate,it can reach 80%.The survival rate of negative control group,CP group and fusion protein group were 20%,30% and 60%,respectively.