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Isolation, Purification, Characterization and Direct Conjugation of the Lipid A-Free Lipopolysaccharide of Vibrio cholerae O139.

Authors
  • Xu, Peng1
  • Korcová, Jana2, 3
  • Baráth, Peter2
  • Čížová, Alžbeta2
  • Valáriková, Jana2
  • Qadri, Firdausi4
  • Kelly, Meagan5
  • O'Connor, Robert D1
  • Ryan, Edward T5, 6, 7
  • Bystrický, Slavomír2
  • Kováč, Pavol1
  • 1 Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, 9000 Rockville, Bethesda, MD, 20892-0815, USA.
  • 2 Institute of Chemistry, Slovak Academy of Sciences, 84238, Bratislava, Slovak Republic.
  • 3 Institute of Epidemiology, Faculty of Medicine, Comenius University, Špitálska 24, 81372, Bratislava, Slovak Republic.
  • 4 Infectious Diseases Division, International Centre for, Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, Bangladesh. , (Bangladesh)
  • 5 Division of Infectious Diseases, Massachusetts General Hospital, Boston, MA, 02114, USA.
  • 6 Harvard Medical School, Boston, MA, 02115, USA.
  • 7 Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Boston, MA, 02115, USA.
Type
Published Article
Journal
Chemistry - A European Journal
Publisher
Wiley (John Wiley & Sons)
Publication Date
Oct 08, 2019
Volume
25
Issue
56
Pages
12946–12956
Identifiers
DOI: 10.1002/chem.201902263
PMID: 31306528
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

The lipopolysaccharide (LPS) of Vibrio cholerae O139, strain CIRS245, was isolated conventionally, and the lipid A was removed by mild acid hydrolysis (0.1 m NaOAc buffer containing 1 % SDS, pH 4.2, 95 °C, 8 h). The crude product was a complex mixture consisting mainly of constituent fragments of the O-specific polysaccharide-core (OSPc). The OSPc was only a minor component in the mixture. Two-stage purification of the crude OSPc by HPLC gave pure OSPc fragment of the LPS, as shown by NMR spectroscopy, analytical HPLC and ESI-MS. This material is the purest OSPc fragment of the LPS from Vibrio cholerae O139 reported to date. The purified OSPc was readily converted to the corresponding methyl squarate derivative and the latter was conjugated to BSA. The conjugate, when examined by ELISA, showed immunoreactivity with sera from patients in Bangladesh recovering from cholera caused by V. cholerae O139, but not O1. © 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

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