Penicillin-resistant monoclonal antibodies and preliminary application of.doc

 PAGE \* MERGEFORMAT 14 Penicillin-resistant monoclonal antibodies and preliminary application of 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 allergy commonly in clinic. METHODS: Penicillin, as a kind of hapten, was conjugated with carrier protein to form complete 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/L.The average recovery rate was 107.81% and the average intra  and inter  coefficient of varitation (CV) was 6.7% and 9.3% respectively. The method was applied for relative quantitative analysis of benzylpenicilloyl protein from Group A Streptococcus Preparation. CONCLUSION: Nine hybridoma cell strains stably secreting mAb against benzylpenicillin were obtained. The double  antibody Sandwich ELISA for relative quantitative analysis of benzylpenicilloyl protein was established. [Keywords] benzylpenicillin; benzylpenicilloyl; mAb; double  antibody Sandwich ELISA [Abstract] Objective: To prepare monoclonal antibodies against penicillin (mAb) and the establishment of double-antibody sandwich ELISA detection methods, the clinical allergic reactions caused by penicillin allergy penicilloyl protein research. Meth