Gene therapy and DNA vaccination are clinical fields gradually emerging in the last few decades, in particular after the discovery of some gene-related diseases. The increased relevance of biomedical applications of plasmid DNA (pDNA) to induce therapeutic effects has had a great impact on biopharmaceutical research and industry. Although there are several steps involved in the pDNA manufacturing process, the several unit operations must be designed and integrated into a global process. After the plasmid has been designed according to the requirements for clinical administeration to humans, it is biosynthesised mainly by an E. coli host. The overriding priority of the production process is to improve plasmid quantity - the production conditions need to be optimised to guarantee pDNA stability and biological activity. The complexity and diversity of biomolecules present on the pDNA-containing extracts represent the main concern and limitation to achieve pure and biologically active pDNA. There has been a recent intenstification of the improvement of existing purification procedures or the establishment of novel schemes for plasmid purification. This review focuses on the progress and relevance of chromatographic methodologies in the purification of pDNA-based therapeutic products. The review will attempt to assemble their different contributions of the different chromatographic procedures that are being used in the pDNA purification area. The advantages and disadvantages of the different chromatographic techniques, as well as the most significant improvements in response to the challenge of purifying pDNA will be discussed, emphasizing the future directions in this field.

中文翻译：

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质粒DNA在基因治疗中的生物医学应用：色谱法的新挑战。

基因治疗和DNA疫苗接种是最近几十年逐渐兴起的临床领域，尤其是在发现一些基因相关疾病之后。质粒DNA（pDNA）在生物医学中诱导治疗效果的相关性日益提高，对生物药物研究和工业产生了重大影响。尽管pDNA制造过程涉及多个步骤，但必须设计多个单元操作并将其集成到全局过程中。根据对人类临床给药的要求设计了质粒后，主要由大肠杆菌宿主生物合成。生产过程的重中之重是提高质粒数量-需要优化生产条件以保证pDNA的稳定性和生物活性。存在于含pDNA的提取物中的生物分子的复杂性和多样性代表了实现纯净的，具有生物活性的pDNA的主要关注点和局限性。最近，对现有纯化方法的改进或用于质粒纯化的新方案的建立已得到加强。这篇综述着重于色谱方法在基于pDNA的治疗产品纯化中的进展和相关性。审查将试图汇总它们在pDNA纯化领域中使用的不同色谱方法的不同贡献。将讨论不同色谱技术的优缺点，以及应对纯化pDNA挑战的最重大改进，