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首页> 外文期刊>Separation and Purification Technology >Anion exchange chromatography of 4.2 kbp plasmid based vaccine (pcDNA3F) from alkaline lysed E. coli lysate using amino functionalised polymethacrylate conical monolith
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Anion exchange chromatography of 4.2 kbp plasmid based vaccine (pcDNA3F) from alkaline lysed E. coli lysate using amino functionalised polymethacrylate conical monolith

机译：使用氨基官能化的聚甲基丙烯酸酯圆锥形整体柱，从碱性裂解的大肠杆菌裂解物中对基于4.2 kbp质粒的疫苗（pcDNA3F）进行阴离子交换层析

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摘要

New strategies to economically manufacture large quantities of vaccines in less time are greatly needed to cater for the increasing global human population. This study aimed at developing a strategy to efficiently manufacture a highly purified plasmid vaccine via anion exchange monolithic chromatography. Diethylamine and triethylamine were used to provide the functional groups for polymethacrylate monoliths and used in anion exchange chromatography of plasmid pcDNA3F. Five chromatographic settings as described in the first table of this article were studied and it was concluded that the triethylamine functionalised conical monolith combined with optimum buffers' and pH conditions produced the highest quality of pcDNA3F. Plasmid yield (3003.55 mg/L), plasmid recovery (90.25%), protein (0.01 mg/L), LPS (0.12 EU/mg) with no detectable gDNA and RNA were obtained at a low NaCl concentration of 0.25 M. Apparently, this technology will have a great impact on the overall plasmid vaccine production and particularly on the development of axial flow monolithic plasmid vaccine purification.
机译：迫切需要在更短的时间内经济地生产大量疫苗的新策略，以适应不断增长的全球人口。这项研究旨在开发一种策略，可以通过阴离子交换整体色谱有效地生产高度纯化的质粒疫苗。二乙胺和三乙胺用于提供聚甲基丙烯酸甲酯整料的官能团，并用于质粒pcDNA3F的阴离子交换色谱中。对本文第一张表中所述的五种色谱设置进行了研究，得出的结论是，三乙胺官能化的锥形整体柱与最佳缓冲液和pH条件相结合可产生最高质量的pcDNA3F。在0.25 M的低NaCl浓度下获得了质粒产量（3003.55 mg / L），质粒回收率（90.25％），蛋白质（0.01 mg / L），LPS（0.12 EU / mg），没有可检测到的gDNA和RNA。该技术将对整个质粒疫苗的生产，特别是对轴流式整体质粒疫苗纯化的发展产生重大影响。

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