High-sensitivity quantification of antisense oligonucleotides for pharmacokinetic characterization.

Aim: Antisense oligonucleotides (ASOs) are a fast-growing drug modality. Pharmacokinetic characterization and accurate quantification of ASOs is critical for drug development. LC-MS and hybridization immunoassays are common methods to quantify ASOs but may lack sensitivity. In this study we aimed to develop an ASO quantification method with improved sensitivity. Methods: We developed a branched DNA approach for ASO quantification and compared it with hybridization immunoassays. Results: The branched DNA assay showed significantly improved sensitivity, with LLOQ 31.25 pg/ml in plasma, 6.4-and 16-fold higher than dual-probe hybridization electrochemiluminescence and single-probe hybridization ELISA, respectively, with adequate precision, accuracy, selectivity and specificity and acceptable matrix interference. Conclusion: Branched DNA for ASO quantification has significantly higher sensitivity and lower hemolysis interference.

Citation

Shilpi Mahajan, Hui Zhao, Kristina Kovacina, Eric Lachacz, Sany Hoxha, Josolyn Chan, Meina Liang. High-sensitivity quantification of antisense oligonucleotides for pharmacokinetic characterization. Bioanalysis. 2022 May;14(9):603-613

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PMID: 35578971

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