Therapeutic oligonucleotides (oligos) have become a focus of interest and promise in biopharmaceutical research due to their potential to treat many kinds of disorders. Whether siRNA, mRNA, or DNA, separating oligos from complex matrix is difficult.

The size, nucleotide complexity, and general variations in these oligos can vary. As a result, liquid chromatography (LC) analysis and purification of oligonucleotides is challenging. We will address chromatographic conditions for oligonucleotide analysis. Specifically, we will discuss fundamental characteristics of the LC column stationary phase, method development guidelines, and optimizations for analytical characterization through scale up to preparative purification.