Objective: To validate the CE-LIF method for N-glycan profile analysis of antibodies inter laboratory. Methods: The method validation was performed according to ICH_Q2_R1 guideline and general chapter 9101 of Chinese Pharmacopoeia (2015 edition). The validation characteristics include 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 content ranging from 100 μg to 400 μg. The R2 of linear regression equation were above 0.97, and the recoveries were between 81% and 109%. For precision test, the RSDs of peak area percentage and retention time of G0F, Man5, G1F(1,6), G1F(1,3) and G2F were below 10%. The quantitation limit of the method was 0.016%, and the range was from 0.016% to 77.233%, whichindicated that single peak at this range could be quantified accurately. Furthermore, robustness evaluation under a series of condition showed that this method was robust, where the RSD of peak area percentage was below 9% and RSD of retention time was below 0.1%. Conclusion: Inter-laboratory method validation of CE-LIF has been organized, and the results verifies that the method is feasible for analysis of antibodies.
WANG Wen-bo, WU Gang, YU Chuan-fei, ZHANG Feng, WANG Lan
. Inter-laboratory method validation of CE-LIF for N-glycan profile analysis of monoclonal antibodies*[J]. Chinese Journal of Pharmaceutical Analysis, 2022
, 42(1)
: 172
-178
.
DOI: 10.16155/j.0254-1793.2022.01.20
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