title: Ultrasensitive Hybridization-Based ELISA Method for the Determination of Phosphorodiamidate Morpholino Oligonucleotides in Biological samples.
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).
authors.affiliation: 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.
authors.forename: Umar
authors.initials: U
authors.lastname: Burki
authors.affiliation: 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.
authors.forename: Volker
authors.initials: V
authors.lastname: Straub
mesh_terms: D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D051379:Mice; D060172:Morpholinos / Q000097:blood* / Q000737:chemistry / Q000235:genetics / Q000493:pharmacokinetics*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D012680:Sensitivity and Specificity; D013223:Statistics as Topic; D014018:Tissue Distribution
chemical_list: D060172:Morpholinos
keywords: Biodistribution; ELISA; Hybridization; MNase; Pharmacokinetic; Phosphorodiamidate morpholino oligonucleotides