Scaleable processes for the manufacture of therapeutic quantities of plasmid DNA
Scaleable processes for the manufacture of therapeutic quantities of plasmid DNA
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用于生产治疗量质粒 DNA 的可扩展工艺
DOI：
10.1042/ba20030011
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发表时间：
2003
影响因子：
2.8
通讯作者：
P. Shamlou
中科院分区：
文献类型：
--
作者：
P. Shamlou
当人们的注意力主要集中在载体设计和早期临床结果的建立上时，生产治疗用质粒DNA(PDNA)的可扩展工艺的需求很容易被忽视。PDNA是一种大分子，其性质类似于受污染的染色体DNA。这些因素，再加上宿主细胞中的低初始质粒浓度，带来了独特的工艺挑战，需要大量的前期设计，以建立稳健的制造工艺，同时也能符合当前的良好制造规范(‘cGMP’)，并生产毫克至千克量的PDNA产品。这篇综述描述了目前正在评估的用于生产治疗性超螺旋PDNA的有前景的可扩展的工艺。综述了提高超螺旋形质粒产量和降低污染物浓度的发酵策略，并评估了下游工艺有效去除细胞污染物的能力，将PDNA的超螺旋形式与其开放的环状和线形分离，并制备用于制剂的纯化药物物质。介绍了目前发展稳定输送系统的策略，并讨论了质量保证和质量控制的方法。
The need for scaleable processes to manufacture therapeutic plasmid DNA (pDNA) is easy to overlook when attention is focused primarily on vector design and establishment of early clinical results. pDNA is a large molecule and has properties that are similar to those of the contaminating chromosomal DNA. These, combined with the low initial concentration of plasmids in the host cell, provide unique process challenges that require significant upfront design to establish robust manufacturing processes that can also comply with current Good Manufacturing Practice (‘cGMP’) and produce milligram‐to‐kilogram quantities of pDNA product. This review describes promising scaleable processes that are currently being assessed for production of therapeutic supercoiled pDNA. Fermentation strategies for improving supercoiled plasmid yield and reducing contaminant concentrations are reviewed, and downstream processes are assessed for their ability to efficiently remove cellular contaminants, separate the supercoiled form of the pDNA from its open circular and linear forms, and prepare the purified drug substance for formulation. Current strategies are presented for developing stable delivery systems, and approaches to quality assurance and quality control are discussed.