Martins R, Queiroz J A, Sousa F
CICS-UBI - Health Sciences Research Centre, University of Beira Interior, Av. Infante D. Henrique, 6200-506 Covilhã, Portugal.
CICS-UBI - Health Sciences Research Centre, University of Beira Interior, Av. Infante D. Henrique, 6200-506 Covilhã, Portugal.
J Chromatogr A. 2014 Aug 15;1355:1-14. doi: 10.1016/j.chroma.2014.05.075. Epub 2014 Jun 6.
Research on RNA has led to many important biological discoveries and improvement of therapeutic technologies. From basic to applied research, many procedures employ pure and intact RNA molecules; however their isolation and purification are critical steps because of the easy degradability of RNA, which can impair chemical stability and biological functionality. The current techniques to isolate and purify RNA molecules still have several limitations and the requirement for new methods able to improve RNA quality to meet regulatory demands is growing. In fact, as basic research improves the understanding of biological roles of RNAs, the biopharmaceutical industry starts to focus on them as a biotherapeutic tools. Chromatographic bioseparation is a high selective unit operation and is the major option in the purification of biological compounds, requiring high purity degree. In addition, its application in biopharmaceutical manufacturing is well established. This paper discusses the importance and the progress of RNA isolation and purification, considering RNA applicability both in research and clinical fields. In particular and in view of the high specificity, affinity chromatography has been recently applied to RNA purification processes. Accordingly, recent chromatographic investigations based on biorecognition phenomena occurring between RNA and amino acids are focused. Histidine and arginine have been used as amino acid ligands, and their ability to isolate different RNA species demonstrated a multipurpose applicability in molecular biology analysis and RNA therapeutics preparation, highlighting the potential contribution of these methods to overcome the challenges of RNA purification.
对RNA的研究已带来许多重要的生物学发现以及治疗技术的改进。从基础研究到应用研究,许多程序都使用纯净且完整的RNA分子;然而,由于RNA易于降解,会损害其化学稳定性和生物学功能,因此其分离和纯化是关键步骤。目前用于分离和纯化RNA分子的技术仍存在一些局限性,并且对能够提高RNA质量以满足监管要求的新方法的需求也在不断增长。事实上,随着基础研究增进了对RNA生物学作用的理解,生物制药行业开始将其作为生物治疗工具加以关注。色谱生物分离是一种高选择性的单元操作,是纯化生物化合物的主要选择,要求具有高纯度。此外,它在生物制药生产中的应用也已得到充分确立。本文讨论了RNA分离和纯化的重要性及进展,同时考虑了RNA在研究和临床领域的适用性。特别是鉴于其高特异性,亲和色谱最近已应用于RNA纯化过程。因此,本文重点介绍了基于RNA与氨基酸之间发生的生物识别现象的最新色谱研究。组氨酸和精氨酸已被用作氨基酸配体,它们分离不同RNA种类的能力在分子生物学分析和RNA治疗制剂制备中显示出多用途适用性,突出了这些方法在克服RNA纯化挑战方面的潜在贡献。