Efficient Method Development of Oligonucleotides by Reversed-Phase Ion-Pair Chromatography

Nucleic acid drugs are produced through chemical synthesis, but their synthesis process often introduces impurities such as shorter and longer lengths of products and protection groups. Therefore, proper chromatographic separation of the target oligonucleotide and impurities is critical for drug quality control and safety investigation. Liquid chromatography (LC) is one of the most popular techniques, combined with reversed-phase ion-pair (RP-IP) chromatography. The separation with RP-IP chromatography is affected by various parameters, including the column chemistry, the concentration of the ion-pair reagent in the mobile phase, organic solvent composition, gradient slope, etc. In addition, the separation performance also varies based on the molecule's chemical properties, such as sequence, length, molecule modifications, etc. Method development of LC separation is obviously challenging when considering the complexity of separation science and molecule chemistry facets. On the other hand, establishing a concrete analytical method in the early phase is vital for lengthy following research work.
To aim to develop a robust LC method for oligonucleotides in a short time, we studied the most efficient method development procedure, from a condition screening through the evaluation of method robustness. In a poster presentation, we will introduce acase of method development with LabSolutions™ MD, designed to support the entire method development procedure based on AQbD concept