Endotoxin detection in a competitive electrochemical assay: Synthesis of a suitable endotoxin conjugate

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A biotin-lipopolysaccharide (biotin-LPS) conjugate was synthesized from LPS smooth from Salmonella minnesota, yielding a conjugate with a biotin/LPS ratio equal to 1:1 and endotoxic activity of 0.08 EU ng-1. The conjugate was used in an amperometric competitive assay to determine endotoxins with end...

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Autores principales:	Priano, G., Pallarola, D., Battaglini, F.
Formato:	JOUR
Materias:	Biotin-conjugated lipopolysaccharide Competitive assay Lipopolysaccharide biotin carboxymethyldextran cystamine endotoxin endotoxin neutralizing protein gold horseradish peroxidase hydrogen peroxide lipopolysaccharide neutravidin unclassified drug amperometry article binding competition chemical modification concentration (parameters) conjugation covalent bond electrochemical analysis electrode molecular recognition nonhuman oxidation reduction reaction priority journal Salmonella minnesota toxin analysis Biotin Electrochemistry Electrodes Endotoxins Lipopolysaccharides Models, Biological Models, Chemical Molecular Structure Oxidation-Reduction Armoracia rusticana Salmonella enterica subsp. enterica serovar Minnesota
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_00032697_v362_n1_p108_Priano
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	todo:paper_00032697_v362_n1_p108_Priano
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spelling	todo:paper_00032697_v362_n1_p108_Priano2023-10-03T13:55:52Z Endotoxin detection in a competitive electrochemical assay: Synthesis of a suitable endotoxin conjugate Priano, G. Pallarola, D. Battaglini, F. Biotin-conjugated lipopolysaccharide Competitive assay Lipopolysaccharide biotin carboxymethyldextran cystamine endotoxin endotoxin neutralizing protein gold horseradish peroxidase hydrogen peroxide lipopolysaccharide neutravidin unclassified drug amperometry article binding competition chemical modification concentration (parameters) conjugation covalent bond electrochemical analysis electrode molecular recognition nonhuman oxidation reduction reaction priority journal Salmonella minnesota toxin analysis Biotin Electrochemistry Electrodes Endotoxins Lipopolysaccharides Models, Biological Models, Chemical Molecular Structure Oxidation-Reduction Armoracia rusticana Salmonella enterica subsp. enterica serovar Minnesota A biotin-lipopolysaccharide (biotin-LPS) conjugate was synthesized from LPS smooth from Salmonella minnesota, yielding a conjugate with a biotin/LPS ratio equal to 1:1 and endotoxic activity of 0.08 EU ng-1. The conjugate was used in an amperometric competitive assay to determine endotoxins with endotoxin-neutralizing protein (ENP) as the recognition element. The assay is performed on a modified electrode, involving the covalent binding of carboxymethyl dextran (CMDex) to a cystamine-modified gold electrode and then the covalent binding of the recognition protein, ENP, to CMDex. The assay is carried out by incubating the modified electrode in an LPS sample to which biotin-LPS was added. Both species compete for the recognition sites on the modified surface. After the incubation stage and a careful rinsing, the electrode is immersed in a solution containing neutravidin-horseradish peroxidase conjugate (N-HRP), which binds to the sites containing biotin-LPS on the electrode. The system is rinsed and a current signal is generated by the addition of hydrogen peroxide and a redox mediator. The assay is able to detect LPS from Salmonella minnesota at concentrations as low as 0.1 ng ml-1, equivalent to 0.07 EU ml-1. © 2006 Elsevier Inc. All rights reserved. Fil:Priano, G. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 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_v362_n1_p108_Priano
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Biotin-conjugated lipopolysaccharide Competitive assay Lipopolysaccharide biotin carboxymethyldextran cystamine endotoxin endotoxin neutralizing protein gold horseradish peroxidase hydrogen peroxide lipopolysaccharide neutravidin unclassified drug amperometry article binding competition chemical modification concentration (parameters) conjugation covalent bond electrochemical analysis electrode molecular recognition nonhuman oxidation reduction reaction priority journal Salmonella minnesota toxin analysis Biotin Electrochemistry Electrodes Endotoxins Lipopolysaccharides Models, Biological Models, Chemical Molecular Structure Oxidation-Reduction Armoracia rusticana Salmonella enterica subsp. enterica serovar Minnesota
spellingShingle	Biotin-conjugated lipopolysaccharide Competitive assay Lipopolysaccharide biotin carboxymethyldextran cystamine endotoxin endotoxin neutralizing protein gold horseradish peroxidase hydrogen peroxide lipopolysaccharide neutravidin unclassified drug amperometry article binding competition chemical modification concentration (parameters) conjugation covalent bond electrochemical analysis electrode molecular recognition nonhuman oxidation reduction reaction priority journal Salmonella minnesota toxin analysis Biotin Electrochemistry Electrodes Endotoxins Lipopolysaccharides Models, Biological Models, Chemical Molecular Structure Oxidation-Reduction Armoracia rusticana Salmonella enterica subsp. enterica serovar Minnesota Priano, G. Pallarola, D. Battaglini, F. Endotoxin detection in a competitive electrochemical assay: Synthesis of a suitable endotoxin conjugate
topic_facet	Biotin-conjugated lipopolysaccharide Competitive assay Lipopolysaccharide biotin carboxymethyldextran cystamine endotoxin endotoxin neutralizing protein gold horseradish peroxidase hydrogen peroxide lipopolysaccharide neutravidin unclassified drug amperometry article binding competition chemical modification concentration (parameters) conjugation covalent bond electrochemical analysis electrode molecular recognition nonhuman oxidation reduction reaction priority journal Salmonella minnesota toxin analysis Biotin Electrochemistry Electrodes Endotoxins Lipopolysaccharides Models, Biological Models, Chemical Molecular Structure Oxidation-Reduction Armoracia rusticana Salmonella enterica subsp. enterica serovar Minnesota
description	A biotin-lipopolysaccharide (biotin-LPS) conjugate was synthesized from LPS smooth from Salmonella minnesota, yielding a conjugate with a biotin/LPS ratio equal to 1:1 and endotoxic activity of 0.08 EU ng-1. The conjugate was used in an amperometric competitive assay to determine endotoxins with endotoxin-neutralizing protein (ENP) as the recognition element. The assay is performed on a modified electrode, involving the covalent binding of carboxymethyl dextran (CMDex) to a cystamine-modified gold electrode and then the covalent binding of the recognition protein, ENP, to CMDex. The assay is carried out by incubating the modified electrode in an LPS sample to which biotin-LPS was added. Both species compete for the recognition sites on the modified surface. After the incubation stage and a careful rinsing, the electrode is immersed in a solution containing neutravidin-horseradish peroxidase conjugate (N-HRP), which binds to the sites containing biotin-LPS on the electrode. The system is rinsed and a current signal is generated by the addition of hydrogen peroxide and a redox mediator. The assay is able to detect LPS from Salmonella minnesota at concentrations as low as 0.1 ng ml-1, equivalent to 0.07 EU ml-1. © 2006 Elsevier Inc. All rights reserved.
format	JOUR
author	Priano, G. Pallarola, D. Battaglini, F.
author_facet	Priano, G. Pallarola, D. Battaglini, F.
author_sort	Priano, G.
title	Endotoxin detection in a competitive electrochemical assay: Synthesis of a suitable endotoxin conjugate
title_short	Endotoxin detection in a competitive electrochemical assay: Synthesis of a suitable endotoxin conjugate
title_full	Endotoxin detection in a competitive electrochemical assay: Synthesis of a suitable endotoxin conjugate
title_fullStr	Endotoxin detection in a competitive electrochemical assay: Synthesis of a suitable endotoxin conjugate
title_full_unstemmed	Endotoxin detection in a competitive electrochemical assay: Synthesis of a suitable endotoxin conjugate
title_sort	endotoxin detection in a competitive electrochemical assay: synthesis of a suitable endotoxin conjugate
url	http://hdl.handle.net/20.500.12110/paper_00032697_v362_n1_p108_Priano
work_keys_str_mv	AT prianog endotoxindetectioninacompetitiveelectrochemicalassaysynthesisofasuitableendotoxinconjugate AT pallarolad endotoxindetectioninacompetitiveelectrochemicalassaysynthesisofasuitableendotoxinconjugate AT battaglinif endotoxindetectioninacompetitiveelectrochemicalassaysynthesisofasuitableendotoxinconjugate
_version_	1807323631391342592

Endotoxin detection in a competitive electrochemical assay: Synthesis of a suitable endotoxin conjugate

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