Feasibility, via a new interface, of mass spectrometry of lipopolysaccharides separated in polyacrylamide gels

Abstract:

For the first time, it was demonstrated the feasibility of analyzing by mass spectrometry (MS) Gram-negative bacteria lipopolysaccharides (LPS) electrophoretically separated with very high resolution in polyacrylamide gels (slab-PAGE). Particularly, the direct impact of this result in four aspects of LPS analysis is demonstrated: spectrometric analysis of the microisolated species, sample preparation for MS, bidimensional separation and heterogeneity of LPS, and the structural characterization of lipopolysaccharides of medical (for example, LPS from strain RM.118 [Rd] of Haemophilus influenzae) and biological interest (for example, LPS from strain HMK of Vibrio fischeri). To build an interface between high-resolution slab-PAGE separation and MS analysis, it was necessary to develop a new method, with unique characteristics, that made possible the microisolation of LPS species: high-efficiency recovery, high precision, no modification of the microisolated material and general applicability (applicable to LPS of different nature and sources). In this way, the bases for the fine structural analysis of the highly complex nature, in molecular species, of bacterial LPS are laid. The experiments showing these results were published in five papers in the leading journals of analytical biochemistry.

Año de publicación:

2005

Keywords:

Fuente:

scopus