High-sensitivity quantification of antisense oligonucleotides for pharmacokinetic characterization.

Disease can be caused by genetic mutations that lead to overproduction or underproduction of an aberrant protein. Antisense oligonucleotides (ASOs) are a relatively new class of drugs. While most current drugs act at the protein level, ASOs work at the RNA level and minimize synthesis of the aberrant protein. ASOs are small synthetic nucleotides that specifically bind and modify the target RNA. Quantification of ASOs is important in drug development to understand how much of the drug is in circulation or in the body after a certain period of time. While there are methods available to quantify ASOs, they lack sensitivity. We developed a method called 'branched DNA' to quantify ASOs, and compared it with known ASO quantification methods. We found that the branched DNA method showed improved sensitivity compared with other existing methods and is a reliable method to quantify ASOs. This method may be used in clinical trials when improved sensitivity quantification is needed and thus facilitate the ASO drug development field.