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Lookup NU author(s): Dr Umar Burki,
Dr Alison Blain,
Dr Steven Laval,
Professor Volker StraubORCiD
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Antisense oligonucleotide (AON)-induced exon skipping is one of the most promising strategies for treating Duchenne muscular dystrophy (DMD) and other rare monogenic conditions. Phosphorodiamidate morpholino oligonucleotides (PMOs) and 2-O-methyl phosphorothioate (2OMe) are two of the most advanced AONs in development. The next generation of peptide-conjugated PMO (P-PMO) is also showing great promise, but to advance these therapies it is essential to determine the pharmacokinetic and biodistribution (PK/BD) profile using a suitable method to detect AON levels in blood and tissue samples. An enzyme-linked immunosorbent assay (ELISA)-based method, which shows greater sensitivity than the liquid chromatography-mass spectrometry method, is the method of choice for 2OMe detection in preclinical and clinical studies. However, no such assay has been developed for PMO/P-PMO detection, and we have, therefore, developed an ultrasensitive hybridization-based ELISA for this purpose. The assay has a linear detection range of 5-250 pM (R-2>0.99) in mouse serum and tissue lysates. The sensitivity was sufficient for determining the 24-h PK/BD profile of PMO and P-PMO injected at standard doses (12.5mg/kg) in mdx mice, the dystrophin-deficient mouse model for DMD. The assay demonstrated an accuracy approaching 100% with precision values under 12%. This provides a powerful cost-effective assay for the purpose of accelerating the development of these emerging therapeutic agents.
Author(s): Burki U, Keane J, Blain A, O'Donovan L, Gait MJ, Laval SH, Straub V
Publication type: Article
Publication status: Published
Journal: Nucleic Acid Therapeutics
Print publication date: 17/09/2015
Online publication date: 15/07/2015
ISSN (print): 2159-3337
ISSN (electronic): 2159-3345
Publisher: Mary Ann Liebert, Inc.
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