细菌内毒素检查法在肺炎球菌结合疫苗研制中的应用及验证

Application and verification of bacterial endotoxin test in preparation of pneumococcal conjugate vaccine

目的应用细菌内毒素检查法(动态浊度法)检测13价肺炎球菌结合疫苗中细菌内毒素含量,并对该方法进行验证。方法用《中国药典》三部(2010版)载录的动态浊度法检查疫苗原辅材料、疫苗过程产物及疫苗细菌内毒素含量,并进行肺炎球菌结合疫苗原液和成品的细菌内毒素含量检测,验证方法的可行性及准确性。结果 13价肺炎球菌结合疫苗生产使用试剂、相关原辅材料和磷酸铝佐剂对细菌内毒素检查无影响;该方法线性、专属性、准确性、精密度、耐用性良好,检定范围初步定为0.08~10 EU/ml,定量限度暂定为0.03 EU/ml;可准确测定疫苗原液和成品中细菌内毒素含量,重复性和准确性良好。结论动态浊度法检测13价肺炎球菌结合疫苗中细菌内毒素含量,结果准确、稳定,符合方法验证的要求,可为该疫苗的质量控制提供稳定可靠的检测方法。

Objective To determine the bacterial endotoxin content in 13-valent pneumococcal conjugate vaccine by bacterial endotoxin test（kinetic turbidimetric assay） and verify the method. Methods Bacterial endotoxin test, i. e.kinetic turbidimetric assay included in Chinese Pharmacopoeia（Volume III, 2010 edition） was used for determination of bacterial endotoxin content in raw and auxiliary materials, intermediate product, bulk, and final product of pneumococcal conjugate vaccine, and verified for feasibility and accuracy. Results The agents, raw and auxiliary materials and aluminium phosphate adjuvant used for production of 13-valent pneumococcal conjugate vaccine showed no impact on bacterial endotoxin test. The method showed good linearity, specificity, accuracy, precision and robustness, of which the detection range was primarily defined as 0. 08 ~ 10 EU/ml while the limit of quantitation as 0. 03 EU/ml. The bacterial endotoxin contents in bulk and final product of vaccine were determined accurately, which indicated good reproducibility and accuracy of the method. Conclusion The determination results of bacterial endotoxin content in 13-valent pneumococcal conjugate vaccine were accurate and stable, which met the requirements for verification of methodology. The study provided a stable and reliable method for quality control of the conjugate vaccine.