Electrophoretic and mass spectrometric strategies for profiling bacterial lipopolysaccharides.
Capillary electrophoresis (CE) is a high-resolution separation technique that has been widely used for trace analysis in biological samples. On-line capillary electrophoresis-electrospray mass spectrometry (CE-MS) was developed for the analysis of lipopolysaccharide (LPS) glycoforms from the gram-negative bacteria, Haemophilus influenzae. In this paper, we report on the application of CE-MS to characterize structural differences in O-deacylated LPS samples from H. influenzae strains Rd 11.7 and 375.1. The resolution capability of on-line CE-MS was first demonstrated by analysis of a complex LPS mixture from H. influenzae strain Rd 11.7. This strain contains a mixture of isomeric glycoforms differing in the number and positions of hexose moieties. Sialic acid containing glycoforms were also determined. Structural features of LPS from a lic1 mutant of H. influenzae strain 375 (375.1) were studied using on-line CE-MS/MS. With the separation provided by CE, two isomeric glycoforms differing in the location of phosphoethanolamine substituents were characterized by tandem mass spectrometry.
Li
Jianjun
J
Institute for Biological Sciences, National Research Council, Ottawa, ON, Canada K1A 0R6. Jianjun.Li@nrc-cnrc.gc.ca
Cox
Andrew D
AD
Hood
Derek W
DW
Schweda
Elke K H
EK
Moxon
E Richard
ER
Richards
James C
JC
D019075:Electrophoresis, Capillary / Q000379:methods; D006193:Haemophilus influenzae / Q000737:chemistry*; D008070:Lipopolysaccharides / Q000032:analysis*; D019158:N-Acetylneuraminic Acid / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization
D008070:Lipopolysaccharides; D019158:N-Acetylneuraminic Acid