Ultrasensitive Hybridization-Based ELISA Method for the Determination of Phosphorodiamidate Morpholino Oligonucleotides in Biological samples

Umar Burki; Volker Straub

doi:10.1007/978-1-4939-6817-6_22

Ultrasensitive Hybridization-Based ELISA Method for the Determination of Phosphorodiamidate Morpholino Oligonucleotides in Biological samples

Methods Mol Biol. 2017:1565:265-277. doi: 10.1007/978-1-4939-6817-6_22.

Authors

Umar Burki¹, Volker Straub²

Affiliations

¹ The John Walton Muscular Dystrophy Research Centre, The MRC Centre for Neuromuscular Diseases, Central Parkway, International Centre for Life, Institute of Genetic Medicine, Newcastle University, Newcastle upon Tyne, NE1 3BZ, UK.
² The John Walton Muscular Dystrophy Research Centre, The MRC Centre for Neuromuscular Diseases, Central Parkway, International Centre for Life, Institute of Genetic Medicine, Newcastle University, Newcastle upon Tyne, NE1 3BZ, UK. volker.straub@ncl.ac.uk.

PMID: 28364250
DOI: 10.1007/978-1-4939-6817-6_22

Abstract

Determining the concentration of oligonucleotide in biological samples such as tissue lysate and serum is essential for determining the biodistribution and pharmacokinetic profile, respectively. ELISA-based assays have shown far greater sensitivities compared to other methods such as HPLC and LC/MS. Here, we describe a novel ultrasensitive hybridization-based ELISA method for quantitating morpholino oligonucleotides in mouse tissue lysate and serum samples. The assay has a linear detection range of 5-250 pM (R2 > 0.99).

Keywords: Biodistribution; ELISA; Hybridization; MNase; Pharmacokinetic; Phosphorodiamidate morpholino oligonucleotides.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Enzyme-Linked Immunosorbent Assay / methods*
Mice
Morpholinos / blood*
Morpholinos / chemistry
Morpholinos / genetics
Morpholinos / pharmacokinetics*
Nucleic Acid Hybridization / methods*
Sensitivity and Specificity
Statistics as Topic
Tissue Distribution

Substances

Morpholinos