【摘要】 目的制备利福霉素单克隆抗体,并建立利福霉素竞争ELISA检测方法。方法通过利福霉素与载体蛋白(BSA、OVA)偶联,制备免疫原和检测抗原,用杂交瘤技术制备单克隆抗体,建立利福霉素竞争ELISA检测方法。结果筛选出5株能稳定分泌抗利福霉素单抗的杂交瘤细胞株,均分泌IgG抗体,其中2H1、5H5、3F12和2C8株的轻链是κ链,3A2株的轻链是λ链;5株单抗为同一位阻群;在碱性条件下较稳定。建立的利福霉素竞争ELISA检测法灵敏度为6ng/ml,与利福平无交叉反应,稳定性良好。结论已成功制备了利福霉素单克隆抗体,并建立了利福霉素竞争ELISA检测法。更多还原

【Abstract】 Objective To prepare the monoclonal antibodies(McAbs)against rifamycin and develop a competitive ELISA method. Methods Rifamycin was coupled to bovine serum albumin(BSA)and ovalbumin(OVA)as carriers respectively. BALB / c mice were immunized with Rifa-BSA to prepare McAbs by hybridoma technique. A competitive ELISA method was developed with the prepared McAbs, using Rifa-OVA as a competitive antigen. Results Five hybridoma cell strains stably secreting McAbs against ri-famycin were screened, and all the secreted McAbs were alkaline-resistant IgG of the same steric group. The light chains of McAbs 2H1, 5H5, 3F12 and 2C8 were κ chains, and that of McAb 3A2 was λ chain. The developed competitive ELISA method showed a sensitivity of 6 ng / ml and good stability, but no cross reaction with rifampin. Conclusion The McAbs against rifamycin were suc-cessfully prepared, and a competitive ELISA method for rifamycin was developed.更多还原