Overcoming Challenges in Oligonucleotide Therapeutics Analysis: A Novel Nonion Pair Approach
Yoshiharu Hayashi1, Yuchen Sun2
1Bioanalysis Research Department, CMIC Pharma Science Co., Ltd., Hyogo 677-0032, Japan.
Abstract:
Oligonucleotide therapeutics (OT) have emerged as promising drug modality for various intractable diseases. Recently, liquid chromatography-mass spectrometry (LC-MS) has been commonly employed for characterizing and quantifying OT in biological samples. Traditionally, the ion pairing-reverse phase (IP-RP) LC-MS method has been utilized in OT bioanalyses; however, this approach is associated with several limitations, including the memory effect and ion suppression effect of IP reagents. Therefore, this study aimed to develop a new RP-LC-MS method that eliminates the need for IP reagents. Our investigation revealed that ammonium bicarbonate was essential for the successful implementation of this nonIP-RP-LC-MS-based bioanalysis of OT. Moreover, the developed method demonstrated high versatility, accommodating the analysis of various natural or chemically modified oligonucleotides. The sensitivity of the method was further assessed using reconstituted plasma samples (the lower limit of quantification in this experiment was 0.5-1 ng/mL). In summary, the developed nonIP-RP-LC-MS method offers an easy, reliable, and cost-effective approach to the bioanalysis of OT.
Insights
A new non-ion pairing, reverse-phase LC-MS method simplifies oligonucleotide therapeutic bioanalysis. This reliable, cost-effective approach overcomes limitations of traditional methods for analyzing these promising drugs.
Area of Science:
- Analytical Chemistry
- Pharmaceutical Sciences
- Biotechnology
Background:
- Oligonucleotide therapeutics (OT) show promise for intractable diseases.
- Liquid chromatography-mass spectrometry (LC-MS) is crucial for OT characterization and quantification.
- Traditional ion-pairing, reverse-phase (IP-RP) LC-MS methods for OT bioanalysis have limitations like memory and ion suppression effects.
Purpose of the Study:
- To develop a novel reverse-phase LC-MS (RP-LC-MS) method for oligonucleotide therapeutic bioanalysis.
- To eliminate the need for ion-pairing reagents in OT bioanalysis.
- To establish a versatile, sensitive, and reliable method for quantifying various oligonucleotides.
Main Methods:
- Development of a non-ion pairing, reverse-phase LC-MS (nonIP-RP-LC-MS) method.
- Utilized ammonium bicarbonate as a key component for successful method implementation.
- Assessed method versatility across natural and chemically modified oligonucleotides; determined sensitivity using reconstituted plasma samples.
Main Results:
- Successfully developed and validated a nonIP-RP-LC-MS method for OT bioanalysis.
- Ammonium bicarbonate was identified as essential for the method's efficacy.
- The method demonstrated high versatility for diverse oligonucleotide types and achieved a lower limit of quantification of 0.5-1 ng/mL in plasma.
Conclusions:
- The developed nonIP-RP-LC-MS method provides an improved approach for OT bioanalysis.
- This method is easy, reliable, and cost-effective, overcoming limitations of traditional IP-RP-LC-MS.
- It offers a versatile platform for the quantification of various oligonucleotide therapeutics in biological matrices.
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