Journal article
Profile of Native N-linked Glycan Structures from Human Serum Using High Performance Liquid Chromatography on a Microfluidic Chip and Time-of-Flight Mass Spectrometry
Proteomics (Weinheim), Vol.9(7), pp.1939-1951
04/2009
Handle:
https://hdl.handle.net/2376/110513
PMCID: PMC2765869
PMID: 19288519
Abstract
Protein glycosylation involves the addition of monosaccharides in a stepwise process requiring no glycan template. Therefore, identifying the numerous glycoforms, including isomers, can help elucidate the biological function(s) of particular glycans. A method to assess the diversity of the N-linked oligosaccharides released from human serum without derivatization has been developed using on-line nano-liquid chromatography (nanoLC) and high resolution time-of-flight mass spectrometry. The N-linked oligosaccharides were analyzed with matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry (MALDI FT-ICR MS) and microchip liquid chromatography mass spectrometry (HPLC-Chip/TOF MS). Two microfluidic chips were employed, the glycan chip (40 nL enrichment column, 43×0.075 mm ID analytical column) and the high capacity chip (160 nL enrichment column, 140×0.075 mm ID analytical column), both with graphitized carbon as the stationary phase. Both chips offered good sensitivity and reproducibility in separating a heterogeneous mixture of neutral and anionic oligosaccharides between injections. Increasing the length and volume of the enrichment and the analytical columns improved resolution of the peaks. Complex type N-linked oligosaccharides were the most abundant oligosaccharides in human serum accounting for ~96% of the total glycans identified, while hybrid and high mannose type oligosaccharides comprise the remaining ~4%.
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Details
- Title
- Profile of Native N-linked Glycan Structures from Human Serum Using High Performance Liquid Chromatography on a Microfluidic Chip and Time-of-Flight Mass Spectrometry
- Creators
- Caroline S Chu - Department of Chemistry, University of California at Davis, Davis, CA 95616Milady R Niñonuevo - Department of Chemistry, University of California at Davis, Davis, CA 95616Brian H Clowers - Department of Chemistry, University of California at Davis, Davis, CA 95616Patrick D Perkins - Agilent Technologies, Inc., Santa Clara, CA 95051Hyun Joo An - Department of Chemistry, University of California at Davis, Davis, CA 95616Hongfeng Yin - Agilent Technologies, Inc., Santa Clara, CA 95051Kevin Killeen - Agilent Technologies, Inc., Santa Clara, CA 95051Suzanne Miyamoto - Division of Hematology and Oncology, University of California Davis Medical Center, Sacramento, CA 95817Rudolf Grimm - Agilent Technologies, Inc., Santa Clara, CA 95051Carlito B Lebrilla - Department of Chemistry, University of California at Davis, Davis, CA 95616
- Publication Details
- Proteomics (Weinheim), Vol.9(7), pp.1939-1951
- Academic Unit
- Chemistry, Department of
- Identifiers
- 99900547761601842
- Language
- English
- Resource Type
- Journal article