Isolation, Purification, Characterization and Direct Conjugation of the Lipid A‐Free Lipopolysaccharide of Vibrio cholerae O139
| Autor: | Meagan Kelly, Firdausi Qadri, Pavol Kováč, Edward T. Ryan, Peng Xu, Robert D. O’Connor, Peter Barath, Jana Valáriková, Alžbeta Čížová, Slavomír Bystrický, Jana Korcová |
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| Rok vydání: | 2019 |
| Předmět: |
Lipopolysaccharides
Spectrometry Mass Electrospray Ionization Magnetic Resonance Spectroscopy Sodium Acetate 010402 general chemistry medicine.disease_cause Polysaccharide 01 natural sciences High-performance liquid chromatography Vibrio cholerae O139 Catalysis Lipid A medicine Chromatography High Pressure Liquid chemistry.chemical_classification Chromatography Strain (chemistry) 010405 organic chemistry Hydrolysis Organic Chemistry General Chemistry Nuclear magnetic resonance spectroscopy bacterial infections and mycoses 0104 chemical sciences chemistry Vibrio cholerae lipids (amino acids peptides and proteins) Acid hydrolysis Conjugate |
| Zdroj: | Chemistry – A European Journal. 25:12946-12956 |
| ISSN: | 1521-3765 0947-6539 |
| DOI: | 10.1002/chem.201902263 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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