SinS - Abstract

Abstract Information :

Oligonucleotides are an increasingly important class of biomolecules that have gained significant interest in recent years due to their therapeutic potential and impact during the covid19 pandemic. However, the complexity and heterogeneity of oligonucleotides along with their physical properties makes the analytical measurement of these molecules challenging. LCMS remains the most commonly used technique for the identification and quantification of oligonucleotides and their impurities. The large size and hydrophilic nature of these molecules make them difficult to retain on traditional reversed-phase columns. Techniques such as ion pair chromatography (IPC) are commonly used but present challenges with regard to toxicity, ion suppression and peak broadening. Hydrophilic interaction chromatography (HILIC) is also commonly used, however, the lack of established methods and the limited availability of suitable stationary phases present their own challenges. The high molecular weight of oligonucleotides, the presence of multiple charged species and the potential for fragmentation further complicates the interpretation of mass spectrometry data. Oligonucleotides are highly hygroscopic, leading to degradation and the chiral nature of the phosphorothioate (P=S) linkage in some oligonucleotides presents further challenge for the separation and analysis of enantiomers. In addition, the lack of International Council for Harmonisation (ICH) guidance for the analysis of oligonucleotides presents a challenge for analytical method development and validation. There is a need for standardised methods and guidelines to ensure the accuracy and reproducibility of results. In short, the importance of these biopharmaceuticals as new medicines and the complexity presented to the pharmaceutical industry in the regulation and analysis of these materials warrants our attention as a community. This presentation will discuss state of the art approaches for the analysis of oligonucelotides from an industrial perspective, highlighting challenges and inviting collaboration on practical solutions to these issues.