Analysis of N-linked Glycan Profile of Therapeutic Antibodies by UPLC-Qda MS
WANG Wen-bo, YU Chuan-fei, ZHANG Feng, XU Gang-ling, GAO Kai, WANG Lan*
Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech Products, National Institutes for Food and Drug Control, Beijing 100050, China
Abstract��OBJECTIVE To analyze the N-glycan profile of therapeutic antibodies by UPLC-Qda MS. METHODS The N-linked glycan in Fc region was released by PNGase F digestion and labeled by RapiFluor-MS, and labeled glycans were analyzed by UPLC-Qda MS. RESULTS The UPLC-Qda MS method showed good accuracy in N-glycan quantitaion and qualification, and the ��m/z value of individual N-glycan was in a range of ��0.5. Three similar biotherapeutic products (SBP) showed a nearly same N-glycan profile with the reference biotherapeutic products (RBP) and the total percentage of fucosylated glycans was comparable, only one fraction of N-glycans had some difference. The major N-glycans of antibodies expressed by CHO cells, NS0 cells and SP2/0 cells were G0F-N, G0F, Man5, G1Fa, G1Fb and G2F, accounting for above 80% of total glycans. Eleven glycoforms were detected in CHO cell expressed antibodies, 22 and 26 glycoforms were detected in NS0 cell and SP2/0 cell expressed antibodies respectively. The N-glycans of NS0 cell and SP2/0 cell expressed antibodies contained more sialylated and galactosylated complex glycoforms, which was related to the antibody half-life in vivo and immunogenicity. CONCLUSION The HILIC UPLC-Qda MS, as a fast and accurate analytical method, can be used in the quality control of N-glycan profile of antibody.
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2018, Vol. 53 
