Oligonucleotides are a growing class of therapeutics used for the treatment of a variety of diseases. A unique aspect of these therapeutics is the variety of technology platforms that can be used to quantify these drugs whether by LC/MS, hELISA, or qPCR. There are many factors that inform the choice of platform to measure oligonucleotides in matrix. In this presentation, we will highlight two hybridization ELISA methods for the quantification of an siRNA. The case study will compare the one-step/S1 nuclease cleavage method and the sandwich hybridization method for quantification of Lumasiran in Rat Plasma. We will highlight important assay parameters such as sensitivity, precision, and accuracy. These results are part of an industry collaboration to compare oligonucleotide quantification methods across technologies. This information will allow researchers to understand the differences in these bioanalytical techniques that can be applied to their drug programs.
Learning Objectives:
Upon completion, participants will be able to gain insights into hELISA methods used for the quantification of siRNA molecules and be able to apply these concepts in their drug programs.
Upon completion, participants will be able to distinguish between a one-step/S1 nuclease and sandwich hybridization assay for siRNA quantification.
Upon completion, participants will be able to evaluate differences in sensitivity, accuracy and precision, and method development between a one-step/S1 nuclease and sandwich hybridization assay for siRNA quantification.
