Validation and Application of Hybridization Liquid Chromatography-Tandem Mass Spectrometry Methods for Quantitative Bioanalysis of Antisense Oligonucleotides

• DOI: 10.4155/bio-2022-0015
• Publication Date: 2022-05-12T09:22:06Z
• Authors: Pei Li; Jean-François Dupuis; Valerie Vrionis; Kevork Mekhssian; Tom Magee; Long Yuan
• Keywords: Tandem Mass Spectrometry; Oligonucleotides; Animals; Reproducibility of Results; Haplorhini; Oligonucleotides, Antisense; 01 natural sciences; Chromatography, Liquid; 0104 chemical sciences

Background: Antisense oligonucleotide (ASO), an emerging modality in drug research and development, demands accurate and sensitive bioanalysis to understand its pharmacokinetic and pharmacodynamic properties. Results: By combining the advantages of both ligand binding and liquid chromatography-mass spectrometry/tandem mass (LC-MS/MS), hybridization LC-MS/MS methods were successfully developed and validated/qualified in a good lab practice (GLP) environment for the quantitation of an ASO drug candidate in monkey serum, cerebrospinal fluid (CSF) and tissues in the range of 0.5-500 ng/ml. Special treatment of CSF samples was employed to mitigate nonspecific binding, improve long-term storage stability and enable the usage of artificial CSF as a more accessible surrogate matrix. The method was also qualified and applied to ASO quantitation in various monkey tissue samples using a cocktail tissue homogenate as a surrogate matrix. Conclusion: This work was the first reported GLP validation and application of ASO bioanalysis using the hybridization LC-MS/MS platform.