Detection of Locked Nucleic Acid Gapmers from Mouse Muscle Samples Using ELISA.

Abstract

Antisense oligonucleotide (ASO)-mediated therapy is promising for the treatment of a variety of genetic disorders, suchas Duchenne muscular dystrophy. As more ASOs advance in therapeutic development and enter clinical trials, it becomes necessary to have a means of quantifying their amounts in biological samples post-treatment. This information will be valuable for evaluating the safety and pharmacokinetic profiles of ASOs, and in deciding how the efficacy of these drugs can be improved. Gapmers are a class of ASOs characterized by having a central DNA portion that is surrounded by chemically modified nucleotides on both ends. While relativelysimpleand accessiblemethods to quantify other ASOs such as phosphorodiamidate morpholino oligomers (PMOs)using enzyme-linked immunosorbent assay (ELISA)-based techniques are available and have been used for in vivo studies, nosuch method is available for gapmers to our knowledge. Here, we describe a sensitive ELISA protocol that can be used to quantify the levels of locked nucleic acid (LNA) gapmers in mouse muscle tissue.