Guide to Reverse Phase HPLC Method Development for Oligonucleotides

Martin Deetz, Nicholas Samulewicz, and Jay Dharia b DuPont

As more companies explore oligonucleotides as part of their biopharma drug pipeline, the importance of developing an effective HPLC method is increasing. While
starting points for mobile phases, ion pairing agent, and example HPLC methods can be found in the literature, often these methods do not transfer across oligos.

In this poster, we use an agile-inspired learning approach to explore gradient and isocratic elution conditions, temperature and flow rate to develop a method
which could provide the oligo in >99% purity and >90% yield. Process development scientists can utilize this methodology and insights to design scalable
processes for high purity oligonucleotides with DuPontb" AmberChromb" XT20 polymeric reverse phase chromatography resin.