In the chromatographic separation process of oligonucleotides (ONs), mechanistic understanding of their binding and diffusion processes is of significant importance to determine operating conditions in a fast and robust way. In this work, we determined the number of binding sites and the diffusivities of ONs in a polymer grafted anion exchange chromatography through linear gradient experiments (LGE) being carried out at selected four to five gradient slopes. Synthetic poly (T)s with length ranging from 3 to 90-mer were employed as a model of an antisense oligonucleotide with typical lengths of 10 - 30 bases. Comparison of the retention was also conducted between the grafted anion exchanger with a conventional ligand and an anion monolith disk. For the ONs up to 50 bases, the number of binding sites determined can be correlated with the length of ONs, and the grafted resin showed a better diffusion and narrower peak width compared to the nongrafted one. The retention behavior became similar for porous media when the longer ONs (> 50mer) were applied. The results obtained suggest that antisense ONs can be separated with grafted ligands without sacrificing mass transfer properties.

在寡核苷酸（ON）的色谱分离过程中，对它们的结合和扩散过程的机械理解对于以快速而稳定的方式确定操作条件非常重要。在这项工作中，我们通过在选定的4至5个梯度斜率上进行的线性梯度实验（LGE），确定了聚合物接枝阴离子交换色谱中结合位点的数量和ON的扩散性。长度为3至90聚体的合成聚（T）被用作典型长度为10至30个碱基的反义寡核苷酸的模型。在具有常规配体的接枝阴离子交换剂和阴离子整体柱之间也进行了保留时间的比较。对于最多50个碱基的ON，所确定的结合位点的数目可以与ON的长度相关，并且与未接枝的树脂相比，接枝的树脂表现出更好的扩散和更窄的峰宽。当使用更长的ON（> 50mer）时，多孔介质的保留行为变得相似。获得的结果表明，可以用嫁接的配体分离反义ON，而不会牺牲传质性能。