Establishing stereochemical comparability in phosphorothioate oligonucleotides with nuclease P1 digestion coupled with LCMS analysis
Zifan Li, Fei Tong, Xiao Li, Nicholas R. Larson, Xuan Zhou, Yueheng Zhang, Jonas P. Immel‐Brown, George M. Bou-Assaf
Abstract
Stereochemical comparability is critical for ensuring manufacturing consistency in therapeutic phosphorothioate oligonucleotides. Currently, analytical methods for this assessment are limited. We hereby report on a novel protocol capable of detecting a stereochemistry change in a single phosphorothioate linkage by employing nuclease P1 digestion of the oligonucleotide with subsequent LCMS analysis of the resulting fragments. The method proves valuable for establishing stereochemical comparability and for ensuring manufacturing consistency of oligonucleotide therapeutics.
Topics & Concepts
ComparabilityOligonucleotideNucleaseChemistryConsistency (knowledge bases)Computational biologyCombinatorial chemistryDNAChromatographyBiochemistryComputer scienceBiologyMathematicsCombinatoricsArtificial intelligenceAdvanced biosensing and bioanalysis techniquesDNA and Nucleic Acid ChemistryRNA Interference and Gene Delivery