Capillary electrophoresis chips for fingerprinting endotoxin chemotypes and subclasses

Béla Kocsis, Lilla Makszin, Anikó Kilár, Zoltán Péterfi, Ferenc Kilár

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Citation (Scopus)

Abstract

Endotoxins (lipopolysaccharides, LPS; lipooligosaccharides, LOS) are components of the envelope of Gram-negative bacteria. These molecules, responsible for both advantageous and harmful biological activity of these microorganisms, are highly immunogenic and directly involved in numerous bacterial diseases in humans, such as Gram-negative sepsis. The characterization of endotoxins is of importance, since their physiological and pathophysiological effects depend on their chemical structure. The differences among the LPS from different bacterial serotypes and their mutants include variations mainly within the composition and length or missing of their O-polysaccharide chains. Microchip electrophoretic methodology enables the structural characterization of LPS molecules from several bacteria and the quantitative evaluation of components of endotoxin extracts. The improved microchip electrophoretic method is based on the direct labeling of endotoxins by covalent binding of a fluorescent dye. The classification of the S-type LPSs can be done according to their electrophoretic profiles, which are characteristics of the respective bacterial strains. According to the number, distribution, and the relative amounts of components in an endotoxin extract, it is possible to differentiate between the S-type endotoxins from different Gram-negative bacterial strains. The microchip electrophoresis affords high-resolution separation of pure and partially purified (e.g., obtained from whole-cell lysate) S and R endotoxins. This microchip technique provides a new, standardizable, fast, and sensitive method for the detection of endotoxins and for the quantitative evaluation of components of an endotoxin extract.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages151-165
Number of pages15
DOIs
Publication statusPublished - Jan 1 2017

Publication series

NameMethods in Molecular Biology
Volume1600
ISSN (Print)1064-3745

Keywords

  • Chemotype
  • Endotoxin
  • Endotoxin profiles
  • Fluorescent labeling
  • LPS
  • Lipopolysaccharide
  • Microchip electrophoresis
  • Whole-cell lysate

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Kocsis, B., Makszin, L., Kilár, A., Péterfi, Z., & Kilár, F. (2017). Capillary electrophoresis chips for fingerprinting endotoxin chemotypes and subclasses. In Methods in Molecular Biology (pp. 151-165). (Methods in Molecular Biology; Vol. 1600). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-6958-6_15