Glycobiology, Vol 9, 335-342, Copyright © 1999 by Oxford University Press
B Monsarrat, T Brando, P Condouret, J Nigou and G Puzo
A new analytical approach based on capillary electrophoresis- electrospray
mass spectrometry (CE/ESI-MS) has provided new insight into the
characterization of mannooligosaccharide caps from lipoarabinomannans
(LAMs), which are key molecules in the immunopathogenesis of tuberculosis.
This analytical approach requires oligosaccharide labeling with the
fluorophore 1-aminopyrene-3,6,8- trisulfonate (APTS) by reductive amination
at the reducing termini. Optimization of the separation and ionization
conditions, such as the choice of capillary electrophoresis (CE)
electrolyte buffers, is presented and discussed. Anionic separation of the
mono and oligosaccharide APTS derivatives was finally achieved with aqueous
triethylammonium formate buffer. It was found that in contrast to the
triethylammonium phosphate buffer, the triethylammonium formate buffer was
appropriate for CE/ESI-MS coupling analysis of APTS-carbohydrate
derivatives. In this case, negative ESI-mass spectra of APTS- carbohydrate
adducts showed mainly (M-2H)2-pseudomolecular ions and some sequence
fragment ions allowing their non-ambiguous structural characterization at
the picomolar level. This analytical approach was successfully applied to
more complex mixtures of carbohydrates released by mild acid hydrolysis of
the lipoarabinomannans from Mycobacterium bovis BCG. The
APTS-mannooligosaccharide cap adducts were separated by CE and their
structural characterization achieved by CE/ESI-MS analyses.
Mannooligosaccharide caps were routinely analyzed by capillary
electrophoresis-laser induced fluorescence (CE-LIF) from 50 fmol of
lipoarabinomannans with mannosyl capping (ManLAMs) but sensitivity was
about 50 times lower using ESI-MS detection.
ORIGINAL ARTICLES
Characterization of mannooligosaccharide caps in mycobacterial lipoarabinomannan by capillary electrophoresis/electrospray mass spectrometry
Institut de Pharmacologie et de Biologie Structurale, UPR 9062 CNRS, 205 Rte de Narbonne, 31077 Toulouse Cedex, France.
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