Abstract
This contribution describes a membrane chromatography purification of a therapeutically useful antisense oligonucleotide (ASO). Sample self-displacement purification conditions were optimized for a 20-mer ASO on a 10-mL bed volume, high-resolution, strong anion-exchange membrane chromatography unit. The scale-up experiments were performed at 177-g scale on a 1-L bed volume unit that resulted in 84.4% yield and 95.0% (analyzed by LC−MS) pure full-length ASO following the membrane chromatography process. The purified ASO was subsequently significantly concentrated and diafiltered into water using a 650-Da MWCO ultrafiltration membrane. This robust, high throughput, membrane-based, anion-exchange chromatographic (AX) method simplifies the process-scale purification of phosphorothioate oligonucleotides by eliminating the need for column packing and packing validation.
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