Development and multicenter validation of an LC–MS-based bioanalytical method for antisense therapeutics

• Published in: Bioanalysis Vol. 14; no. 18; pp. 1213 - 1227
• Main Authors: Sun, Yuchen, Nitta, Shin-ichiro, Saito, Kosuke, Hosogai, Ryuta, Nakai, Keiko, Goda, Ryoya, Shimizu, Hisao, Fujita, Hisashi, Kakehi, Masaaki, Murata, Kazuyuki, Yamaguchi, Takeru, Okuzono, Takeshi, Yamane, Shinichi, Kawabata, Mitsuhiko, Matsunuma, Takayuki, Takahara, Kentaro, Kato, Noriko, Yamada, Masaki, Yoshida, Tokuyuki, Inoue, Takao, Saito, Yoshiro
• Format: Journal Article
• Language: English
• Published: Newlands Press Ltd 01-09-2022
• Subjects: antisense oligonucleotide; Clarity OTX; ion-pairing reagent; LC–MS bioanalysis; multilaboratory validation
• ISSN: 1757-6180; 1757-6199
• DOI: 10.4155/bio-2022-0126

Many bioanalytical methods for antisense oligonucleotides (ASOs) using LC–MS have been reported. However, no data have been available on the reproducibility and robustness of a single bioanalytical method for ASOs. As such, in the current study, we evaluated the reproducibility and robustness of LC–MS-based bioanalytical methods for ASOs in multiple laboratories.
Seven independent laboratories were included in this study. Mipomersen was measured by ion-pairing LC–MS (IP-LC–MS) as a model ASO using different LC–MS. The validation results of calibration curve, accuracy, precision and selectivity met the criteria of conventional bioanalytical method validation guidelines using LC/GC–MS for drugs in all laboratories. Meanwhile, carryover (>20%) was detected in three laboratories.
We first demonstrated the multicenter-validated IP-LC–MS bioanalytical method for ASOs. Our data showed that the method was sensitive, robust and reproducible. However, the occurrence of carryover should be carefully monitored in its future application.