Simultaneous quantification of siRNA antisense and sense strands by hybrid liquid chromatography-mass spectrometry.

ABSTRACT:

Background

Most oligonucleotide bioanalytical assays currently only quantify the pharmacologically-active antisense strand, though there have been recent efforts to simultaneously quantify the sense strand using hybridization ELISA or solid phase extraction LC-MS. Hybrid LC-MS, which offers both high sensitivity and specificity unlike the currently used platforms, has not been applied to quantify both siRNA strands simultaneously.

Materials & methods

A hybrid LC-MS assay utilizing LNA capture probes was developed and applied to quantify both strands of a 21-mer lipid-conjugated siRNA (SIR-3) using tandem mass spectrometry (MS/MS). A similar approach using high-resolution mass spectrometry (HRMS) was also evaluated.

Results

The final LC-MS/MS method was capable of quan

SUBMITTER: Agrawal K 

PROVIDER: S-EPMC11864311 | biostudies-literature | 2025 Feb

REPOSITORIES: biostudies-literature