Purification of single-stranded oligonucleotides using anion exchange chromatography
Date
March 21, 2022
The production of pure single-stranded oligonucleotides requires efficient, scalable and cost-effective chromatographic purification methods applied after chemical synthesis. Key impurities in crude samples are length-based and are best removed by anion exchange chromatography. In this case study, we utilized a high-performance strong anion exchange chromatography resin for purification as demonstrated with two oligonucleotides (20-mer and 21-mer) having a phosphodiester backbone. Key process parameters like flow rate, gradient slope, buffer pH and dynamic binding capacity were explored and optimized. Both yield (~90%) and purity (~98%) data were obtained during scale- up studies. In addition, this study indicated that the strong anion exchange resin can also be used for the efficient purification of fully thiolated oligonucleotides used in clinical trials.
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