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

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posted on 2022-11-21, 10:10 authored by Yuchen Sun, Shin-ichiro Nitta, Kosuke Saito, Ryuta Hosogai, Keiko Nakai, Ryoya Goda, Hisao Shimizu, Hisashi Fujita, Masaaki Kakehi, Kazuyuki Murata, Takeru Yamaguchi, Takeshi Okuzono, Shinichi Yamane, Mitsuhiko Kawabata, Takayuki Matsunuma, Kentaro Takahara, Noriko Kato, Masaki Yamada, Tokuyuki Yoshida, Takao Inoue, Yoshiro Saito

Background: 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. Methods/Results:

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.

Conclusion: 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.


This work was supported by the Japan Agency for Medical Research and Development (AMED) (grant nos. JP17-21AK0101073 and JP22AK0101185j0101). S Nitta, R Hosogai and K Nakai are employees of LSI Medience Corporation. R Goda is an employee of Daiichi Sankyo Company Ltd. H Shimizu, H Fujita and M Kakehi are employees of Takeda Pharmaceutical Company Ltd. K Murata and T Yamaguchi are employees of Sumika Chemical Analysis Service Ltd. T Okuzono and S Yamane are employees of Sekisui Medical Co. Ltd. M Kawabata is an employee of Shin Nippon Biomedical Laboratories Ltd. T Matsunuma and K Takahara are employees of Thermo Fisher Scientific KK. N Kato and M Yamada are employees of Shimadzu Corporation.