Development of a bioanalytical method for an antisense therapeutic using high-resolution mass spectrometry

• Published in: Bioanalysis Vol. 12; no. 24; pp. 1739 - 1756
• Main Authors: Sun, Yuchen, Nitta, Shin-ichiro, Saito, Kosuke, Hosogai, Ryuta, Nakai, Keiko, Goda, Ryoya, Kakehi, Masaaki, Murata, Kazuyuki, Yamaguchi, Takeru, Okuzono, Takeshi, Yamane, Shinichi, Enoki, Yoshimasa, Kawabata, Mitsuhiko, Takahara, Kentaro, Sato, Shimpei, Yoshida, Tokuyuki, Inoue, Takao, Saito, Yoshiro
• Format: Journal Article
• Language: English
• Published: London Newlands Press Ltd 01-12-2020; Newlands Press
• Subjects: antisense oligonucleotide; Antisense oligonucleotides; Antisense therapy; Calibration; Chromatography; clarity OTX; Deoxyribonucleic acid; DNA; Gene expression; high-resolution mass spectrometry; Hybridization; ion-pairing reagent; liquid chromatography; Mass spectroscopy; Metabolism; Metabolites; Methods; Plasma; Quality control; United States > US; Japan
• ISSN: 1757-6180; 1757-6199
• DOI: 10.4155/bio-2020-0225

Ion-pairing reverse-phase LC coupled with high-resolution mass spectrometry (IP-LC/HRMS) has gained attention in oligonucleotide therapeutic bioanalyses owing to its high sensitivity and selectivity. However, optimization and validation of IP-LC/HRMS-based methods are rare. The objective of this study is the development of a sensitive and reproducible IP-LC/HRMS-based bioanalytical method using clinically approved mipomersen as a model for antisense oligonucleotides.
Mipomersen was extracted from rat plasma using Clarity OTX SPE and quantified by IP-LC/HRMS. The calibration range was 0.5–250.0 ng/ml. The developed method met the general regulatory criteria for accuracy, precision, carry-over, selectivity, matrix effect and dilution integrity.
A highly sensitive and reliable method for mipomersen measurement with potential antisense oligonucleotide bioanalysis applications has been developed.