Endotoxin detection in a competitive electrochemical assay: Synthesis of a suitable endotoxin conjugate
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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2007
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paper:paper_00032697_v362_n1_p108_Priano2023-06-08T14:24:04Z Endotoxin detection in a competitive electrochemical assay: Synthesis of a suitable endotoxin conjugate Priano, Graciela Inés Pallarola, Diego Andrés Battaglini, Fernando 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. 2007 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00032697_v362_n1_p108_Priano 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, Graciela Inés Pallarola, Diego Andrés Battaglini, Fernando 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. |
| author |
Priano, Graciela Inés Pallarola, Diego Andrés Battaglini, Fernando |
| author_facet |
Priano, Graciela Inés Pallarola, Diego Andrés Battaglini, Fernando |
| author_sort |
Priano, Graciela Inés |
| 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 |
| publishDate |
2007 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00032697_v362_n1_p108_Priano http://hdl.handle.net/20.500.12110/paper_00032697_v362_n1_p108_Priano |
| work_keys_str_mv |
AT prianogracielaines endotoxindetectioninacompetitiveelectrochemicalassaysynthesisofasuitableendotoxinconjugate AT pallaroladiegoandres endotoxindetectioninacompetitiveelectrochemicalassaysynthesisofasuitableendotoxinconjugate AT battaglinifernando endotoxindetectioninacompetitiveelectrochemicalassaysynthesisofasuitableendotoxinconjugate |
| _version_ |
1768544343717576704 |