Interlaboratory Validation of HILIC-UPLC Method for N-glycan Profile Analysis of Monoclonal Antibodies / 中国药学杂志

ABSTRACT

OBJECTIVE: To validate the HILIC-UPLC method for N-glycan profile analysis of therapeutic antibodies. METHODS: The interlaboratory method validation was performed according to ICH_Q2_R1 guideline and general principles of China Pharmacopoeia (2015 edition) 9101. The validation items included specificity, linearity, accuracy, precision, quantitation limit, range and robustness. RESULTS: The method showed good specificity, accuracy, precision and robustness, and showed a good linearity at a protein range from 100 to 400 μg. The r2 of linear regression equation was above 0.98, and the recoveries were between 86% and 117%. Both the RSDs of peak area percentage and retention time were below 10%, which indicated good precision. The lower quantitation limit of the method was 0.040%, and the range was from 0.040% to 78.751%, which means that single peak at this range could be quantified accurately. Furthermore, robustness evaluation under a series of conditions showed that this method was robust, where the RSD of peak area percentage was below 5% and RSD of retention time was below 1%. CONCLUSION: Interlaboratory validation of HILIC-UPLC provides a methodological verification basis for the improvement of Chinese Pharmacopoeia standards.