Extraction of Oligonucleotides From Plasma Samples Across Multiple Species Using OligoWorks™ SPE Microplate Kit

Oligonucleotide therapeutics are a key focus area for many drug developers today given their powerful ability to address disease biology at the level of gene transcription and translation with high target specificity and low toxicity. They are designed with specific types of modifications that enhance protein binding, half-life, and cellular uptake. Given their diversity, developing robust bioanalytical methods for oligonucleotide analysis can be quite challenging, and sample preparation is often its most challenging aspect due to the tight oligonucleotide-to-protein binding that occurs in biomatrices, often leading to low extraction recoveries and poor sensitivity. This work demonstrates use of the OligoWorks SPE Microplate Kit with RapiZyme™ Proteinase K Digestion to effectively extract oligonucleotides from plasma samples across multiple preclinical animal species including human. Using the starting protocol with no adjustments, oligonucleotide recoveries between 56–115% were obtained.