Synthetic oligonucleotide therapeutics provide an effective treatment for a variety of devastating diseases. Within the biopharmaceutical industry, the request for purification and analytical techniques has increased to meet the demands of expanded pipelines. In this study, we demonstrate the effectiveness of a one-step hydrophobic interaction chromatography (HIC) for the purification of 5′-DMT (4,4'-dimethoxytrityl)-on oligonucleotides. The low pH on-column DMT cleavage method results in high DMT-off oligonucleotide recovery while maintaining >95% product purity. Additionally, we detail analytical methods including reversed-phase chromatography, analytical anion-exchange chromatography, and size exclusion chromatography, to ensure the high purity and quality of the final product.
