抗青霉素单克隆抗体的制备及初步应用.doc

抗青霉素单克隆抗体的制备及初步应用
Preparation and primary application of monoclonal antibody against benzylpenicillin



[Abstract] AIM: To develop monoclonal antibody(mAb) against benzylpenicillin and to establish a Sandwich ELISA method for relative quantitative analysis of the allergen which induced penicillin monly in clinic. METHODS: Penicillin, as a kind of hapten, was conjugated with carrier protein to plete antigen and then was used to immunize BALB/c mice. Hybridoma cells secrecting mAb against penicillin were developed. Ascites from the immunized mice were purified by caprylic acid ammonium sulfate precipitation. The specificity of the purified antibodies was detected and the suitable antibodies were used for establishing Sandwich ELISA. RESULTS: Nine hybridoma cells stably secrecting mAb were prepared through cell fusion, screening and cloning, and five of these purified mAb had relative high affinity. The doubleantibody Sandwich ELISA for relative quantitative analysis of benzylpenicilloyl protein was established with a sensitivity of 870U/ average recovery rate was %
and the average intra and intercoefficient of varitation (CV) was % and % respectively. The method was applied for relative quantitative analysis of benzylpenicilloyl protein from Group A us Preparation. CONCLUSION: Nine hybridoma cell strains stably secreting mAb against benzylpenicil