Scientific Director Altasciences Laval, Quebec, Canada
Therapeutic oligonucleotides, such as antisense oligonucleotides (ASOs) and small interfering RNA (siRNA), are an increasingly important class of drugs in the field of drug discovery and development. Accurate measurement of these oligonucleotides in biological samples is crucial for studying their pharmacokinetic and pharmacodynamic properties. Recently, hybridization LC-MS/MS has emerged as a highly sensitive and specific technique for quantifying single- and double-stranded therapeutic oligos. This method demonstrates exceptional specificity by distinguishing the target oligonucleotide from closely related sequences. The hybridization step, which utilizes full complementary DNA or PNA probes, ensures a high level of sensitivity, comparable to ligand-binding assays. This work provides an overview of the characteristics and benefits of the hybridization LC-MS/MS workflow, emphasizing the key differences in its application for ASOs and siRNA compounds. The approach is applicable to different biological matrices and can be combined with different microsampling techniques, allowing the collection of low sample volumes ( < 20uL).
Learning Objectives:
Define the characteristics and advantages of the hybridization LC-MS/MS workflow for the quantification of therapeutic oligonucleotides in biological matrices
Understand the differences in the hybridization approach between single-stranded (ASOs) and double-stranded (siRNA) oligonucleotides
Explore novel applications of hybridization LC-MS workflow including its coupling to microsampling sample collection techniques