抗青霉素单克隆抗体的制备及初步应用
青霉素;,青霉噻唑基;,单克隆抗体;,双抗体夹心ELISA,,]青霉素;,青霉噻唑基;,单克隆抗体;,双抗体夹心ELISA,1材料和方法,2结果,3讨论,参考文献:
Preparation and primary application of monoclonal antibody against benzylpenicillinWU Yu, MENG Shufang, HU Changqin, JIN Shaohong*
National Institude for the Control of Pharmaceutical and Biological Products, Beijing 100050, China
[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 doubleantibody 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 intercoefficient 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 doubleantibody Sandwich ELISA for relative quantitative analysis of benzylpenicilloyl protein was established. ......
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