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Isolation, Purification, Characterization and Direct Conjugation of the Lipid A-Free Lipopolysaccharide of Vibrio cholerae O139.
Chemistry 2019C

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.

Authors+Show Affiliations

Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, 9000 Rockville, Bethesda, MD, 20892-0815, USA.Institute of Chemistry, Slovak Academy of Sciences, 84238, Bratislava, Slovak Republic. Institute of Epidemiology, Faculty of Medicine, Comenius University, Špitálska 24, 81372, Bratislava, Slovak Republic.Institute of Chemistry, Slovak Academy of Sciences, 84238, Bratislava, Slovak Republic.Institute of Chemistry, Slovak Academy of Sciences, 84238, Bratislava, Slovak Republic.Institute of Chemistry, Slovak Academy of Sciences, 84238, Bratislava, Slovak Republic.Infectious Diseases Division, International Centre for, Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, Bangladesh.Division of Infectious Diseases, Massachusetts General Hospital, Boston, MA, 02114, USA.Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, 9000 Rockville, Bethesda, MD, 20892-0815, USA.Division of Infectious Diseases, Massachusetts General Hospital, Boston, MA, 02114, USA. Harvard Medical School, Boston, MA, 02115, USA. Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Boston, MA, 02115, USA.Institute of Chemistry, Slovak Academy of Sciences, 84238, Bratislava, Slovak Republic.Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, 9000 Rockville, Bethesda, MD, 20892-0815, USA.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31306528

Citation

Xu, Peng, et al. "Isolation, Purification, Characterization and Direct Conjugation of the Lipid A-Free Lipopolysaccharide of Vibrio Cholerae O139." Chemistry (Weinheim an Der Bergstrasse, Germany), 2019.
Xu P, Korcová J, Baráth P, et al. Isolation, Purification, Characterization and Direct Conjugation of the Lipid A-Free Lipopolysaccharide of Vibrio cholerae O139. Chemistry. 2019.
Xu, P., Korcová, J., Baráth, P., Čížová, A., Valáriková, J., Qadri, F., ... Kováč, P. (2019). Isolation, Purification, Characterization and Direct Conjugation of the Lipid A-Free Lipopolysaccharide of Vibrio cholerae O139. Chemistry (Weinheim an Der Bergstrasse, Germany), doi:10.1002/chem.201902263.
Xu P, et al. Isolation, Purification, Characterization and Direct Conjugation of the Lipid A-Free Lipopolysaccharide of Vibrio Cholerae O139. Chemistry. 2019 Jul 15; PubMed PMID: 31306528.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Isolation, Purification, Characterization and Direct Conjugation of the Lipid A-Free Lipopolysaccharide of Vibrio cholerae O139. AU - Xu,Peng, AU - Korcová,Jana, AU - Baráth,Peter, AU - Čížová,Alžbeta, AU - Valáriková,Jana, AU - Qadri,Firdausi, AU - Kelly,Meagan, AU - O'Connor,Robert D, AU - Ryan,Edward T, AU - Bystrický,Slavomír, AU - Kováč,Pavol, Y1 - 2019/07/15/ PY - 2019/05/31/received PY - 2019/07/09/revised PY - 2019/7/16/pubmed PY - 2019/7/16/medline PY - 2019/7/16/entrez KW - NMR spectroscopy KW - Vibrio cholerae KW - glycoconjugate vaccines KW - polysaccharides KW - squaric acid JF - Chemistry (Weinheim an der Bergstrasse, Germany) JO - Chemistry N2 - 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. SN - 1521-3765 UR - https://www.unboundmedicine.com/medline/citation/31306528/Isolation,_purification,_characterization_and_direct_conjugation_of_the_lipid_A-free_lipopolysaccharide_of_Vibrio_cholerae_O139 L2 - https://doi.org/10.1002/chem.201902263 DB - PRIME DP - Unbound Medicine ER -