Journal
JOURNAL OF CHROMATOGRAPHY A
Volume 1600, Issue -, Pages 105-111Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2019.04.059
Keywords
N-glycans; Glycosylamine labeling; Filter-aided extraction; HILIC-HPLC/FLD
Funding
- National Natural Science Foundation of China [20905027]
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Efficient sample pretreatment of N-glycans from glycoproteins is essential but challenging due to the limitations of existing tedious and laborious methods in N-glycomics. This study aimed to establish a filter-aided extraction method coupled with glycosylamine AQC labeling for a simple and rapid direct HPLC-FLD-based analysis of N-glycans. The developed method was demonstrated to be simpler and more sensitive compared to previous HILIC SPE purification method coupled with glycosylamine labeling. It has been validated with wild-type N-glycans from human transferrin and RNase B and then was successfully applied to investigate N-glycan profiles of the transferrin in human serum and a monoclonal antibody (mAb). Results showed good applicability of the method for complex samples. Additionally, this method is compatible with the replicate determination of N-glycan samples to assess the high-throughput analysis of glycan variability in mAb sample. (C) 2019 Elsevier B.V. All rights reserved.
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