Sensitive and specific quantification of antisense oligonucleotides using probe alteration-linked self-assembly reaction technology

Quantitative bioanalysis is essential when establishing pharmacokinetic properties during the drug development process. To overcome challenges of sensitivity, specificity and process complexity associated with the conventional analysis of antisense oligonucleotides (ASOs), a new approach to nonenzym...

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Main Authors: Masako Osawa, Takurou Akiya, Funa Ogawa, Takao Suzuki, Masaki Yamagami, Tadashi Umemoto, Akira Ideno
Format: Article
Language:English
Published: Taylor & Francis Group 2023-07-01
Series:BioTechniques
Subjects:
Online Access:https://www.future-science.com/doi/10.2144/btn-2023-0005
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author Masako Osawa
Takurou Akiya
Funa Ogawa
Takao Suzuki
Masaki Yamagami
Tadashi Umemoto
Akira Ideno
author_facet Masako Osawa
Takurou Akiya
Funa Ogawa
Takao Suzuki
Masaki Yamagami
Tadashi Umemoto
Akira Ideno
author_sort Masako Osawa
collection DOAJ
description Quantitative bioanalysis is essential when establishing pharmacokinetic properties during the drug development process. To overcome challenges of sensitivity, specificity and process complexity associated with the conventional analysis of antisense oligonucleotides (ASOs), a new approach to nonenzymatic hybridization assays using probe alteration-linked self-assembly reaction (PALSAR) technology as a signal amplifier was evaluated. PALSAR quantification of ASOs in mouse tissue and plasma was able to achieve a high sensitivity ranging from 1.5 to 6 pg/ml, intra-/interday accuracies in the range of 86.8–119.1% and 88.1–113.1%, respectively, and precision of ≤17.2%. Furthermore, crossreactivity of 3′n-1, a metabolite with a single base difference, was <1%. Our approach provides an auspicious method for distinguishing metabolites and detecting ASOs with high sensitivity and specificity.
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spelling doaj.art-c59bd1e7c8ef452f9e11b7123f8f64852024-11-02T04:16:36ZengTaylor & Francis GroupBioTechniques0736-62051940-98182023-07-01751213010.2144/btn-2023-0005Sensitive and specific quantification of antisense oligonucleotides using probe alteration-linked self-assembly reaction technologyMasako Osawa0Takurou Akiya1Funa Ogawa2Takao Suzuki3Masaki Yamagami4Tadashi Umemoto5Akira Ideno61New Technology, Tsukuba Research Institute, Research &amp; Development, Sekisui Medical Co., Ltd, Inashiki-gun, Ibaraki, 300-1155, Japan1New Technology, Tsukuba Research Institute, Research &amp; Development, Sekisui Medical Co., Ltd, Inashiki-gun, Ibaraki, 300-1155, Japan1New Technology, Tsukuba Research Institute, Research &amp; Development, Sekisui Medical Co., Ltd, Inashiki-gun, Ibaraki, 300-1155, Japan2Luxna Biotech Co., Ltd, Suita, Osaka, 565-0871, Japan2Luxna Biotech Co., Ltd, Suita, Osaka, 565-0871, Japan2Luxna Biotech Co., Ltd, Suita, Osaka, 565-0871, Japan1New Technology, Tsukuba Research Institute, Research &amp; Development, Sekisui Medical Co., Ltd, Inashiki-gun, Ibaraki, 300-1155, JapanQuantitative bioanalysis is essential when establishing pharmacokinetic properties during the drug development process. To overcome challenges of sensitivity, specificity and process complexity associated with the conventional analysis of antisense oligonucleotides (ASOs), a new approach to nonenzymatic hybridization assays using probe alteration-linked self-assembly reaction (PALSAR) technology as a signal amplifier was evaluated. PALSAR quantification of ASOs in mouse tissue and plasma was able to achieve a high sensitivity ranging from 1.5 to 6 pg/ml, intra-/interday accuracies in the range of 86.8–119.1% and 88.1–113.1%, respectively, and precision of ≤17.2%. Furthermore, crossreactivity of 3′n-1, a metabolite with a single base difference, was <1%. Our approach provides an auspicious method for distinguishing metabolites and detecting ASOs with high sensitivity and specificity.https://www.future-science.com/doi/10.2144/btn-2023-0005antisense oligonucleotidebioanalysisligand-binding assaynonenzymatic hybridization assayPALSAR technologysensitivity
spellingShingle Masako Osawa
Takurou Akiya
Funa Ogawa
Takao Suzuki
Masaki Yamagami
Tadashi Umemoto
Akira Ideno
Sensitive and specific quantification of antisense oligonucleotides using probe alteration-linked self-assembly reaction technology
BioTechniques
antisense oligonucleotide
bioanalysis
ligand-binding assay
nonenzymatic hybridization assay
PALSAR technology
sensitivity
title Sensitive and specific quantification of antisense oligonucleotides using probe alteration-linked self-assembly reaction technology
title_full Sensitive and specific quantification of antisense oligonucleotides using probe alteration-linked self-assembly reaction technology
title_fullStr Sensitive and specific quantification of antisense oligonucleotides using probe alteration-linked self-assembly reaction technology
title_full_unstemmed Sensitive and specific quantification of antisense oligonucleotides using probe alteration-linked self-assembly reaction technology
title_short Sensitive and specific quantification of antisense oligonucleotides using probe alteration-linked self-assembly reaction technology
title_sort sensitive and specific quantification of antisense oligonucleotides using probe alteration linked self assembly reaction technology
topic antisense oligonucleotide
bioanalysis
ligand-binding assay
nonenzymatic hybridization assay
PALSAR technology
sensitivity
url https://www.future-science.com/doi/10.2144/btn-2023-0005
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