Székely László, Kiessig Steffen, Schwarz Maria A, Kálmán Franka
Protein Analytics-Development, Solvias AG, Basel, Switzerland.
Electrophoresis. 2009 May;30(9):1579-86. doi: 10.1002/elps.200800380.
Recently, several therapeutic double-stranded (ds) oligonucleotides (ODNs) are in pharmaceutical development. During quality control, these therapeutic molecules have to be characterized with respect to their identity, their content and their impurity profile. It follows that the ds molecule as well as its process- and product-related impurities have to be quantified. The single strands are considered as process as well as product-related impurities in the ds drug substance. Applying well known, conventional, single-base resolution CE-CGE systems developed for the quality control of single-stranded antisense ODNs in the early 1990s, it turned out that the ds ODNs under investigation are migrating in broad, splitted peaks between the peaks reaction zones are observed. It follows that the quantification of the single strands in the drug substance as well as quantification of other product-related impurities, e.g. n-1; n-2 (loss of one and two bases (n), respectively) etc., are not possible without adaptation of the test system. The paper shows how the test system was adjusted in order to determine single-stranded strands as well as ds strands next to each other quantitatively in the ds drug substance under investigation.
最近，几种治疗性双链（ds）寡核苷酸（ODN）正处于药物研发阶段。在质量控制过程中，这些治疗性分子必须在其身份、含量和杂质概况方面进行表征。因此，ds分子及其与工艺和产品相关的杂质都必须进行定量分析。单链在ds原料药中被视为与工艺和产品相关的杂质。应用20世纪90年代初为单链反义ODN质量控制开发的众所周知的传统单碱基分辨率CE-CGE系统，结果发现所研究的ds ODN在宽的、分裂的峰中迁移，在峰反应区之间观察到。因此，如果不调整测试系统，就无法对原料药中的单链进行定量分析，也无法对其他与产品相关的杂质进行定量分析，例如n-1；n-2（分别为一个和两个碱基（n）的缺失）等。本文展示了如何调整测试系统，以便在被研究的ds原料药中同时定量测定单链和ds链。
