Maximizing Oligonucleotide Yield Without Sacrificing Purity

Purifying Antisense Oligonucleotides (ASOs) via traditional single-column batch chromatography often leaves downstream teams facing a frustrating trade-off: sacrifice valuable target product in impure side-fractions, or settle for lower overall purity.

Our latest application note highlights how switching from conventional batch methodology to Multicolumn Countercurrent Solvent Gradient Purification (MCSGP) continuous twin-column technology overcomes this yield-purity tradeoff.

By utilizing MCSGP with AutoPeak® technology, impure product-containing side-fractions are automatically recycled inline in a periodic closed-loop process. The result? Maximum product recovery without accumulating impurities.

Key insights from the study comparing batch vs. MCSGP on BioPro IEX SmartSep Q20 resin:
► 1.8x Yield Increase: Scaled on the Contichrom TWIN system, MCSGP increased recovery yield from 47.9% to 84.3% while boosting purity to 95.8%.
► Seamless Scaling: Successful translation of method development from benchtop (Contichrom CUBE) directly to pilot/production scale (Contichrom TWIN).
► Consistent High Purity: Both small and pilot-scale MCSGP runs reliably purified raw DNA 20mer ASO crude (from ~72% starting purity) to >94% final purity.

A special thank you to our collaborators at PeptiStar, Inc. for their valuable insights and contributions to this work!

Download the Application Note.