Design and optimization of a chromatographic purification process for Streptococcus pneumoniae serotype 23F capsular polysaccharide by a Design of Experiments approach
Design and optimization of a chromatographic purification process for Streptococcus pneumoniae serotype 23F capsular polysaccharide by a Design of Experiments approach
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DOI：
10.1016/j.chroma.2014.04.096
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发表时间：
2014-06-27
影响因子：
4.1
通讯作者：
Sui, Lili
中科院分区：
文献类型：
--
作者：
Ji, Yu;Tian, Yang;Sui, Lili
多价肺炎球菌疫苗在世界范围内用于保护人类免受肺炎球菌疾病的侵害。为了消除传统疫苗纯化工艺中使用的有毒有机溶液，本研究提出了肺炎链球菌血清型23 F荚膜多糖（CPS）的替代色谱工艺。将实验设计策略引入到工艺开发中，解决了工艺开发过程中复杂的设计过程。通过初步的工艺分析，对整个流程进行了回顾，并通过FMEA确定了色谱分离的关键因素，根据原料的性质选择了流通模式。然后进行树脂筛选研究以选择候选树脂。利用DoE生成分辨率IV部分析因设计，以进一步比较候选项并缩小设计空间。选择Capto Adhere后，执行Box-Behnken DoE以模拟该过程并表征因素对响应的所有影响。最后，采用蒙特卡罗模拟对工艺进行优化，对所选的最佳工艺条件进行检验，确定控制限。在设定值下的三次放大运行结果验证了DoE和模拟预测。最终结果完全符合欧盟药典要求：蛋白质/CPS（w/w）1.08%; DNA/CPS（w/w）0.61%;磷含量3.1%;氮0.315%和甲基戊糖百分比47.9%。最终纯CPS的其他检测也符合药典质量标准。这种不含有毒有机溶剂的肺炎球菌疫苗的替代色谱纯化工艺是通过DoE方法成功开发的，并证明了可扩展性、耐用性和大规模生产的适用性。(C)2014爱思唯尔有限公司版权所有。
Multivalent pneumococcal vaccines were used worldwide to protect human beings from pneumococcal diseases. In order to eliminate the toxic organic solutions used in the traditional vaccine purification process, an alternative chromatographic process for Streptococcus pneumoniae serotype 23F capsular polysaccharide (CPS) was proposed in this study. The strategy of Design of Experiments (DoE) was introduced into the process development to solve the complicated design procedure. An initial process analysis was given to review the whole flowchart, identify the critical factors of chromatography through FMEA and chose the flowthrough mode due to the property of the feed. A resin screening study was then followed to select candidate resins. DoE was utilized to generate a resolution IV fractional factorial design to further compare candidates and narrow down the design space. After Capto Adhere was selected, the Box-Behnken DoE was executed to model the process and characterize all effects of factors on the responses. Finally, Monte Carlo simulation was used to optimize the process, test the chosen optimal conditions and define the control limit. The results of three scale-up runs at set points verified the DoE and simulation predictions. The final results were well in accordance with the EU pharmacopeia requirements: Protein/CPS (w/w) 1.08%; DNA/CPS (w/w) 0.61%; the phosphorus content 3.1%; the nitrogen 0.315% and the Methyl-pentose percentage 47.9%. Other tests of final pure CPS also met the pharmacopeia specifications. This alternative chromatographic purification process for pneumococcal vaccine without toxic organic solvents was successfully developed by the DoE approach and proved scalability, robustness and suitability for large scale manufacturing. (C) 2014 Elsevier B.V. All rights reserved.