CICS-UBI - Health Sciences Research Centre, University of Beira Interior, Covilhã, Portugal.
CDRSP-IPLEIRIA - Centre for Rapid and Sustainable Product Development, Instituto Politécnico de Leiria, Marinha Grande, Portugal.
Methods Mol Biol. 2022;2466:135-144. doi: 10.1007/978-1-0716-2176-9_10.
Nucleic acid-based therapy has been emerging as a new strategy with great potential for the treatment of numerous diseases, especially those caused by gene defects. In this context, biotechnology plays a critical role on establishing suitable processes for biopharmaceuticals manufacturing, while the purification step still imposes a major burden. Affinity chromatography using amino acids as specific ligands has been successfully applied for plasmid DNA purification. In this protocol, we describe the process for nucleic acids production and extraction, as well as the chromatographic matrix synthesis for separation between DNA and RNA. This novel arginine-macroporous support presents excellent binding capacity and great robustness for nucleic acids isolation.
核酸治疗作为一种新兴策略，在治疗多种疾病方面具有巨大的潜力，特别是由基因缺陷引起的疾病。在这种情况下，生物技术在建立适合生物制药制造的工艺方面发挥着关键作用，而纯化步骤仍然是一个主要的负担。使用氨基酸作为特定配体的亲和层析已成功应用于质粒 DNA 的纯化。在本方案中，我们描述了核酸的生产和提取过程，以及用于分离 DNA 和 RNA 的色谱基质合成过程。这种新型的精氨酸大孔载体具有出色的核酸结合能力和较强的稳定性。
