Sinha Nanda D, Jung Kyeong Eun
Independent Consultant, Boxborough, Massachusetts.
ST Pharm Co. Ltd, Gyeonggi-do, South Korea.
Curr Protoc Nucleic Acid Chem. 2015 Jun 3;61:10.5.1-10.5.39. doi: 10.1002/0471142700.nc1005s61.
Chromatographic methods have been essential tools for analysis and purification of synthetic oligonucleotides since the 1970s. Significant developments in terms of instruments and stationary phases (media) have been made during the past several decades; among the latest are sub-micron to micron particles for the media, as well as ultra performance liquid chromatography (UPLC). Micron and sub-micron particles have increased product resolution. Applications of recently developed methods such as IP-RP-HPLC and LC-MS have been discussed for analysis, along with use of various methods for purification. Utilization of UPLC has decreased analysis time, increasing the throughput for analysis. Commonly used methods for analysis and purification of synthetic oligonucleotides have been described in this unit.
自20世纪70年代以来,色谱方法一直是分析和纯化合成寡核苷酸的重要工具。在过去几十年中,仪器和固定相(介质)方面取得了重大进展;最新的进展包括用于介质的亚微米到微米级颗粒,以及超高效液相色谱(UPLC)。微米级和亚微米级颗粒提高了产物分辨率。本文讨论了诸如离子对反相高效液相色谱(IP-RP-HPLC)和液相色谱-质谱联用(LC-MS)等最新开发方法在分析中的应用,以及各种纯化方法的使用。UPLC的应用缩短了分析时间,提高了分析通量。本单元介绍了合成寡核苷酸分析和纯化的常用方法。