Ligand-binding assays vs chromatographic platforms for oligonucleotide quantification
Amy White
Author for correspondence editor@bioanalysisjournal.com
First draft submitted: 24 February 2023; Accepted for publication: 24 February 2023; Published online:
6 March 2023
Keywords: antisense oligonucleotides drug development EUSALBAS LC-MS therapeutics
Oligonucleotides are, in essence, nucleic acid polymers that have the potential to treat a vast array of diseases, acting as therapeutics that predominantly focus on gene silencing to. Their typical mechanism of action involves binding of the oligonucleotide to a target mRNA sequence, hindering the action of ribosomes, disrupting splicing and encouraging the action of enzymes that degrade the mRNA. Oligonucleotides can be chemically modified to increase their potency. For instance, the addition a fluorine molecule to the 2′ ribose position increases both the binding affinity of the oligonucleotide and its resilience to nuclease proteins
The two most widely used strategies for gene silencing employ antisense oligonucleotides (ASOS) and small interfering RNAs (siRNAs). There are many different oligonucleotide therapeutic modalities but all aid in targeting specific diseases that are currently untreatable by existing therapies. In general, oligonucleotide therapeutics, such as mRNA or noncoding RNA, have high specificity and can target molecules that cannot be governed by standard drugs, thus resulting in the cultivation of ground-breaking drugs against genetic diseases and cancers beneficial for future treatment of patients.
Due to their many benefits, the interest in oligonucleotide therapeutics has increased over the years, thus the consideration for their analysis, regulation and validation must be a key focus moving forward. The current method of validation-oligonucleotide quantification is an integral step in ensuring drug ufery and efficacy by evaluating oligonucleotide concentration. Ligand-binding (LBA) and LC-MS platforms are the most preferred platforms used for quantification of therapeutic modalities. The essence of a LBA is that a ligand is used to capeate and/or detect the target molecule typically large molecules within biological matrices) and, due to its high specificity, can differentiate Laga molecules better than chromatographic methods. The process of LC-MS analysis corails Liquid chromatography (1) separation wachined with MS in order to detect contacted analyses, in this instance euclides, al matrices and due to specificity and dynamic range, can provide
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