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