Enhancement of fucosylated N-glycan isomer separation with an ultrahigh column temperature in porous graphitic carbon liquid chromatography-mass spectrometry
| Autor: | Jung-Lee Lin, Chung-Hsuan Chen, Nan-Hon Lin, Chung-Yi Wu, Chein-Hung Chen, Sachin S. Shivatare, Chien-Tai Ren, Ya-Ping Lin |
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| Rok vydání: | 2020 |
| Předmět: |
Glycan
Glycosylation Time Factors 010402 general chemistry Tandem mass spectrometry Mass spectrometry 01 natural sciences Biochemistry Analytical Chemistry Ion chemistry.chemical_compound Hydrolysis Isomerism Liquid chromatography–mass spectrometry Polysaccharides Tandem Mass Spectrometry Derivatization Fucose Ions Chromatography biology 010401 analytical chemistry Organic Chemistry Temperature Dextrans General Medicine 0104 chemical sciences Dextran chemistry biology.protein Graphite Porosity Chromatography Liquid |
| Zdroj: | Journal of chromatography. A. 1632 |
| ISSN: | 1873-3778 |
| Popis: | Due to the heterogeneous and isomeric nature of glycans, the development of an advanced separation of distinct glycan isomers is essential for glycan research and application. In this study, we utilized porous graphite carbon (PGC) chromatography for the separation of isomeric oligosaccharides without reduction or chemical derivatization at 190 °C in a custom-built heating oven. Furthermore, the fine structures of glycan isomers could be identified by using ultrahigh temperature PGC liquid chromatography mass spectrometry (UHT-PGC-LCMS). A nonreduced hydrolyzed dextran was applied to verify the performance of UHT-PGC. When the temperature of the PGC column was increased from 25 to 190 °C, the liquid chromatography separation power of the nonreduced dextran ladder significantly increased. The advantage of the UHT-PGC column was its high peak capacity with gradient elution in 10 min at 190 °C, 6700 psi, and a 250 μL/min flow rate for native glycan analysis. Four synthetic Lewis antigen isomers were used to elucidate the separation effectiveness in UHT-PGC. Moreover, mass spectrometry-based sequencing to generate specific diagnostic ions from the four synthetic Lewis antigens was used to predict isomeric glycans based on the relative intensity ratio (RIR) of diagnostic ions. The intensities of the diagnostic ions of synthetic isomers were used to identify each isomer of the fucosylated glycan. The results clearly showed that terminal Lewis A and X residues were in the 3- and 6-arms of N-glycan, respectively. |
| Databáze: | OpenAIRE |
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