【作者】 刘英微； 王曦； 柳蔷； 何亚丽； 张德有； 马锐； 李津；

【Author】 LIU Ying-wei1,WANG Xi1,LIU Qiang2,HE Ya-li2,ZHANG De-you1,MA Rui1,LI Jin(1,3) (1.Vaccine Research DepartmentⅡ,Beijing Tiantan Biological Products Co.,Ltd.,Beijing 100024,China; 2.HB Vaccine Department,Beijing Tiantan Biological Products Co.,Ltd.; 3.Microbiology Department,Peking University Health Science Center)

【机构】 北京天坛生物制品股份有限公司 疫苗研究二室； 北京天坛生物制品股份有限公司乙肝疫苗室； 北京大学医学部基础医学院病毒生物学系；

【摘要】 目的:检测重组汉逊酵母表达HBsAg各纯化工序样品中内毒素含量,分析各工序对内毒素的去除作用。方法:采用鲎试剂法检测各批各纯化HBsAg工序样品中内毒素含量,计算出各工序的内毒素去除率,并分析各纯化工序与内毒素去除率的关系。结果:通过检测各批次的纯化工序样品中内毒素含量和蛋白含量,计算出内毒素去除率。内毒素含量由细胞破碎样品的265 EU/mL下降至最后工序样品的1.0 EU/mL以下。结论:纯化工序微滤、超滤和疏水层析可有效去除内毒素。更多还原

【Abstract】 Objective:By detecting the endotoxin content in the sample of each purification process of HBsAg derived from recombinant Hansenula polymorpha to evaluate the endotoxin removal efficacy of each purification steps.Methods: LAL assay was used to detect the endotoxin content in the sample of each HBsAg purification process for different purification runs.Results:By detecting endotoxin content of the sample of every purification steps in each run,the endotoxin removal rate of each step was calculated.The endotoxin content in samples decreased from the 265 EU/mL in cell disruption step to the 1.0 EU/mL in final sterile filtration.Conclusion:The purifying stages of microfiltration,ultrafiltration and hydrophobic chromatography can remove endotoxin effectively during the purification process of recombinant Hansenula polymorpha derived HBsAg.更多还原