PMID- 10222001
OWN - NLM
STAT- MEDLINE
DCOM- 19990615
LR  - 20061115
IS  - 0003-2697 (Print)
IS  - 0003-2697 (Linking)
VI  - 269
IP  - 2
DP  - 1999 May 1
TI  - Analysis of carbohydrate heterogeneity in a glycoprotein using liquid
      chromatography/mass spectrometry and liquid chromatography with tandem mass
      spectrometry.
PG  - 297-303
AB  - High-performance liquid chromatography with on-line electrospray ionization mass 
      spectrometry (ESI-LC/MS) was investigated for the analysis of carbohydrate
      heterogeneity using RNase B as a model glycoprotein. Oligosaccharides released
      from RNase B with endoglycosidase H were reduced and separated on a graphitized
      carbon column (GCC). GCC-HPLC/MS in the positive-ion mode was successful in the
      identification of one Man5GlcNAc, three Man6GlcNAc, three Man7GlcNAc, three
      Man8GlcNAc, one Man9GlcNAc, and an oligosaccharide having six hexose units (Hex) 
      and two N-acetylhexosamine units (HexNAc). The branch structures of the three
      Man7GlcNAc isomers were determined by liquid chromatography with tandem mass
      spectrometry (LC/MS/MS). LC/MS/MS analysis was shown to be useful for the
      detection and identification of a trace amount of Hex6HexNAc2 alditol as a
      hybrid-type oligosaccharide. Its structure was confirmed by the combination of
      LC/MS with enzymatic digestion using beta-galactosidase and
      N-acetyl-beta-glucosaminidase. The relative quantities of high-mannose-type
      oligosaccharides in RNase B detected by ESI-LC/MS are in reasonable agreement
      with those by UV, high-pH anion-exchange chromatography with pulsed amperometric 
      detection, fluorophore-assisted carbohydrate electrophoresis. Our results
      indicate that LC/MS and LC/MS/MS can be utilized to elucidate the distribution of
      oligosaccharides and their structures, which differ in molecular weight, sugar
      sequence, and branch structure.
CI  - Copyright 1999 Academic Press.
FAU - Kawasaki, N
AU  - Kawasaki N
AD  - Division of Biological Chemistry and Biologicals, National Institute of Health
      Sciences, 1-18-1 Kamiyoga, Tokyo, Setagaya-ku, 158-8501, Japan.
FAU - Ohta, M
AU  - Ohta M
FAU - Hyuga, S
AU  - Hyuga S
FAU - Hashimoto, O
AU  - Hashimoto O
FAU - Hayakawa, T
AU  - Hayakawa T
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Anal Biochem
JT  - Analytical biochemistry
JID - 0370535
RN  - 0 (Carbohydrates)
RN  - 0 (Glycoproteins)
RN  - 0 (Oligosaccharides)
RN  - 0 (Sugar Alcohols)
RN  - EC 3.1.- (Ribonucleases)
RN  - EC 3.1.27.- (ribonuclease B)
SB  - IM
MH  - Animals
MH  - Carbohydrate Sequence
MH  - Carbohydrates/*analysis/chemistry
MH  - Cattle
MH  - Chromatography, High Pressure Liquid/*methods
MH  - Glycoproteins/*chemistry
MH  - Mass Spectrometry/*methods
MH  - Molecular Sequence Data
MH  - Oligosaccharides/analysis/chemistry
MH  - Ribonucleases/chemistry
MH  - Sugar Alcohols/analysis
EDAT- 1999/05/01 00:00
MHDA- 1999/05/01 00:01
CRDT- 1999/05/01 00:00
PHST- 1999/05/01 00:00 [pubmed]
PHST- 1999/05/01 00:01 [medline]
PHST- 1999/05/01 00:00 [entrez]
AID - S0003-2697(99)94026-5 [pii]
AID - 10.1006/abio.1999.4026 [doi]
PST - ppublish
SO  - Anal Biochem. 1999 May 1;269(2):297-303. doi: 10.1006/abio.1999.4026.