Oligonucleotides are polymeric sequences of nucleotides (RNA, DNA, and their analogs) that are utilized extensively as PCR and microarray-based reagents in life science research and DNA-based diagnostic test kits (target specific primers and probes). Increasingly they are being developed as therapeutic drugs for a wide range of disease conditions. While there are just eight FDA-approved oligonucleotide-based therapeutic drugs on the market today, there are over 200 in the clinical pipeline and many more in earlier stages of development.
Oligonucleotide, or nucleic acid-based therapeutics hold great promise and are being developed across a wide range of modalities – from antisense oligonucleotides (AON) and small interfering RNA (siRNA) molecules that make up the majority of the clinical pipeline today, to microRNA inhibitors (antiMir’s) and activators (ProMir’s), aptamers, and most recently, CRISPR-based drugs for gene editing as well as full-length messenger RNA transcripts (mRNA) for gene therapy and vaccine applications.
To enhance their stability in-vivo, nucleic acid-based therapeutics often incorporate chemically-modified nucleotides, and increasingly are being conjugated to other chemical moieties (GalNac, Cholesterol, Peptides, Antibodies, Fatty Acids, etc.), or are being encapsulated within lipid nano-particles or viral vectors (e.g.: Adeno-Associated Virus (AAV) particles) in order to achieve targeted delivery to specific cells within the body.
Like all other biologics, the biophysical characteristics and purity of these molecules must be precisely understood and controlled to meet regulatory requirements. Most oligonucleotides are produced via an automated solid-phase synthesis process, which invariably results in failure sequences and other process-related impurities that need to be separated to the greatest extent possible, and impurities as low as 0.005% of the full-length product (FLP) that co-elute or closely elute with the FLP must also be identified and fully characterized to ensure the safety and efficacy of the final drug formulation.
By combining our ACQUITY UPLC instrument technology with our industry leading Oligo Separation Technology (OST) columns, and pre-validated MassPrep OST reference standard, Waters brings an unrivaled level of chromatographic performance for oligonucleotide separations and analysis.
To this front-end separation, we offer several fit-for-purpose mass analysis options on the back-end to meet the needs of our customers:
Of these four workflows, three utilize our MassLynx Software for instrument control and data acquisition, and leverage ProMass for Masslynx (Novatia, Ltd.) to carry out automated data analysis and reporting. Our new BioAccord mass confirmation workflow utilizes UNIFI providing a single complete audit trail for acquisition, processing, and reporting, reducing risk and simplifying audit preparation.
To learn more about these workflows and the performance benefits that they provide, please have a look at the following:
Oligonucleotide Applications Notebook