An efficient method for conjugation of a lipopolysaccharide from Salmonella enterica sv. Minnesota with probes bearing hydrazine or amino functional groups

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A conjugation method for coupling probes bearing hydrazine or primary amino groups to a lipopolysaccharide (LPS) is described. LPS is modified through the hydroxyl groups present in its O-antigen moiety by activation with cyanogen bromide in aqueous acetone using triethylamine to enhance the electro...

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Autores principales:	Pallarola, D., Battaglini, F.
Formato:	JOUR
Materias:	Cyanogen Bromide Dansyl hydrazine Horseradish peroxidase Lipopolysaccharide conjugates amino acid horseradish peroxidase hydrazine lipid A lipopolysaccharide article bacterium conjugation biological activity death labeling index multiple organ failure nonhuman priority journal Salmonella enterica shock toxicity Amines Chemistry, Analytical Dansyl Compounds Electrophoresis Horseradish Peroxidase Hydrazines Molecular Probes Polyethylene Glycols Polysaccharides, Bacterial Spectrometry, Fluorescence Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Armoracia rusticana
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_00032697_v381_n1_p53_Pallarola
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	todo:paper_00032697_v381_n1_p53_Pallarola
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spelling	todo:paper_00032697_v381_n1_p53_Pallarola2023-10-03T13:55:52Z An efficient method for conjugation of a lipopolysaccharide from Salmonella enterica sv. Minnesota with probes bearing hydrazine or amino functional groups Pallarola, D. Battaglini, F. Cyanogen Bromide Dansyl hydrazine Horseradish peroxidase Lipopolysaccharide conjugates amino acid horseradish peroxidase hydrazine lipid A lipopolysaccharide article bacterium conjugation biological activity death labeling index multiple organ failure nonhuman priority journal Salmonella enterica shock toxicity Amines Chemistry, Analytical Dansyl Compounds Electrophoresis Horseradish Peroxidase Hydrazines Molecular Probes Polyethylene Glycols Polysaccharides, Bacterial Salmonella enterica Spectrometry, Fluorescence Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Armoracia rusticana Salmonella enterica A conjugation method for coupling probes bearing hydrazine or primary amino groups to a lipopolysaccharide (LPS) is described. LPS is modified through the hydroxyl groups present in its O-antigen moiety by activation with cyanogen bromide in aqueous acetone using triethylamine to enhance the electrophilicity of CNBr. The method yields conjugates with good labeling ratios, preserving the endotoxic activity of the lipid A moiety, which in blood exerts pleiotropic effects on many tissues and organs, resulting in multiple-organ damage, circulatory collapse, and death. Conjugation of smooth-form LPS from Salmonella enterica sv. Minnesota to dansyl hydrazine yielded a labeling ratio of 330 nmol dansyl/mg LPS, with nearly no loss of the original endotoxic activity. In the case of horseradish peroxidase, in which a spacer was introduced, the ratio was 28 nmol HRP/mg of LPS, preserving 65% of the original endotoxic activity. This work shows that under these conditions of CNBr activation, the labeling process has practically no effect on the endotoxic behavior of LPS. The method can be used effectively for the conjugation of LPS to probes bearing primary amino, hydrazine, or hydrazide functional groups. © 2008 Elsevier Inc. All rights reserved. Fil:Pallarola, D. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Battaglini, F. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00032697_v381_n1_p53_Pallarola
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Cyanogen Bromide Dansyl hydrazine Horseradish peroxidase Lipopolysaccharide conjugates amino acid horseradish peroxidase hydrazine lipid A lipopolysaccharide article bacterium conjugation biological activity death labeling index multiple organ failure nonhuman priority journal Salmonella enterica shock toxicity Amines Chemistry, Analytical Dansyl Compounds Electrophoresis Horseradish Peroxidase Hydrazines Molecular Probes Polyethylene Glycols Polysaccharides, Bacterial Salmonella enterica Spectrometry, Fluorescence Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Armoracia rusticana Salmonella enterica
spellingShingle	Cyanogen Bromide Dansyl hydrazine Horseradish peroxidase Lipopolysaccharide conjugates amino acid horseradish peroxidase hydrazine lipid A lipopolysaccharide article bacterium conjugation biological activity death labeling index multiple organ failure nonhuman priority journal Salmonella enterica shock toxicity Amines Chemistry, Analytical Dansyl Compounds Electrophoresis Horseradish Peroxidase Hydrazines Molecular Probes Polyethylene Glycols Polysaccharides, Bacterial Salmonella enterica Spectrometry, Fluorescence Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Armoracia rusticana Salmonella enterica Pallarola, D. Battaglini, F. An efficient method for conjugation of a lipopolysaccharide from Salmonella enterica sv. Minnesota with probes bearing hydrazine or amino functional groups
topic_facet	Cyanogen Bromide Dansyl hydrazine Horseradish peroxidase Lipopolysaccharide conjugates amino acid horseradish peroxidase hydrazine lipid A lipopolysaccharide article bacterium conjugation biological activity death labeling index multiple organ failure nonhuman priority journal Salmonella enterica shock toxicity Amines Chemistry, Analytical Dansyl Compounds Electrophoresis Horseradish Peroxidase Hydrazines Molecular Probes Polyethylene Glycols Polysaccharides, Bacterial Salmonella enterica Spectrometry, Fluorescence Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Armoracia rusticana Salmonella enterica
description	A conjugation method for coupling probes bearing hydrazine or primary amino groups to a lipopolysaccharide (LPS) is described. LPS is modified through the hydroxyl groups present in its O-antigen moiety by activation with cyanogen bromide in aqueous acetone using triethylamine to enhance the electrophilicity of CNBr. The method yields conjugates with good labeling ratios, preserving the endotoxic activity of the lipid A moiety, which in blood exerts pleiotropic effects on many tissues and organs, resulting in multiple-organ damage, circulatory collapse, and death. Conjugation of smooth-form LPS from Salmonella enterica sv. Minnesota to dansyl hydrazine yielded a labeling ratio of 330 nmol dansyl/mg LPS, with nearly no loss of the original endotoxic activity. In the case of horseradish peroxidase, in which a spacer was introduced, the ratio was 28 nmol HRP/mg of LPS, preserving 65% of the original endotoxic activity. This work shows that under these conditions of CNBr activation, the labeling process has practically no effect on the endotoxic behavior of LPS. The method can be used effectively for the conjugation of LPS to probes bearing primary amino, hydrazine, or hydrazide functional groups. © 2008 Elsevier Inc. All rights reserved.
format	JOUR
author	Pallarola, D. Battaglini, F.
author_facet	Pallarola, D. Battaglini, F.
author_sort	Pallarola, D.
title	An efficient method for conjugation of a lipopolysaccharide from Salmonella enterica sv. Minnesota with probes bearing hydrazine or amino functional groups
title_short	An efficient method for conjugation of a lipopolysaccharide from Salmonella enterica sv. Minnesota with probes bearing hydrazine or amino functional groups
title_full	An efficient method for conjugation of a lipopolysaccharide from Salmonella enterica sv. Minnesota with probes bearing hydrazine or amino functional groups
title_fullStr	An efficient method for conjugation of a lipopolysaccharide from Salmonella enterica sv. Minnesota with probes bearing hydrazine or amino functional groups
title_full_unstemmed	An efficient method for conjugation of a lipopolysaccharide from Salmonella enterica sv. Minnesota with probes bearing hydrazine or amino functional groups
title_sort	efficient method for conjugation of a lipopolysaccharide from salmonella enterica sv. minnesota with probes bearing hydrazine or amino functional groups
url	http://hdl.handle.net/20.500.12110/paper_00032697_v381_n1_p53_Pallarola
work_keys_str_mv	AT pallarolad anefficientmethodforconjugationofalipopolysaccharidefromsalmonellaentericasvminnesotawithprobesbearinghydrazineoraminofunctionalgroups AT battaglinif anefficientmethodforconjugationofalipopolysaccharidefromsalmonellaentericasvminnesotawithprobesbearinghydrazineoraminofunctionalgroups AT pallarolad efficientmethodforconjugationofalipopolysaccharidefromsalmonellaentericasvminnesotawithprobesbearinghydrazineoraminofunctionalgroups AT battaglinif efficientmethodforconjugationofalipopolysaccharidefromsalmonellaentericasvminnesotawithprobesbearinghydrazineoraminofunctionalgroups
_version_	1807323219313557504

An efficient method for conjugation of a lipopolysaccharide from Salmonella enterica sv. Minnesota with probes bearing hydrazine or amino functional groups

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