An alternative method for the detection of endotoxins

F. BATTAGLINI; G. PRIANO; D. PALLAROLA

San Antonio, Texas

Congreso; Joint meeting of the Society for Leukocyte Biology and the International Endotoxin and Innate Immunity Society; 2006

International Endotoxin Society

During the last twenty years, a great improvement has been made in the study of chemical interactions between lipopolysaccharides and antibacterial agents produced by many organisms. These findings allow the development of an assay equivalent to those based upon antigen-antibody interaction, where the selective recognition is combined with a physicochemical change, which is used as a transducer element generating a signal proportional to the concentration of the analyte to be detected. We propose a competitive electrochemical assay using a modified gold electrode with a recombinant endotoxin neutralizing protein (ENP) from Saccharomyces cerevisiae as recognition element. Two strategies were used to immobilize the protein onto the electrode, one based on electrostatic interactions (electrostatic configuration) and the other trough covalent binding (covalent configuration). LPS from S. minnesota and E. coli were tested in a competitive assay with LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support strategies were used to immobilize the protein onto the electrode, one based on electrostatic interactions (electrostatic configuration) and the other trough covalent binding (covalent configuration). LPS from S. minnesota and E. coli were tested in a competitive assay with LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support strategies were used to immobilize the protein onto the electrode, one based on electrostatic interactions (electrostatic configuration) and the other trough covalent binding (covalent configuration). LPS from S. minnesota and E. coli were tested in a competitive assay with LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support proportional to the concentration of the analyte to be detected. We propose a competitive electrochemical assay using a modified gold electrode with a recombinant endotoxin neutralizing protein (ENP) from Saccharomyces cerevisiae as recognition element. Two strategies were used to immobilize the protein onto the electrode, one based on electrostatic interactions (electrostatic configuration) and the other trough covalent binding (covalent configuration). LPS from S. minnesota and E. coli were tested in a competitive assay with LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support strategies were used to immobilize the protein onto the electrode, one based on electrostatic interactions (electrostatic configuration) and the other trough covalent binding (covalent configuration). LPS from S. minnesota and E. coli were tested in a competitive assay with LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support strategies were used to immobilize the protein onto the electrode, one based on electrostatic interactions (electrostatic configuration) and the other trough covalent binding (covalent configuration). LPS from S. minnesota and E. coli were tested in a competitive assay with LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support proportional to the concentration of the analyte to be detected. We propose a competitive electrochemical assay using a modified gold electrode with a recombinant endotoxin neutralizing protein (ENP) from Saccharomyces cerevisiae as recognition element. Two strategies were used to immobilize the protein onto the electrode, one based on electrostatic interactions (electrostatic configuration) and the other trough covalent binding (covalent configuration). LPS from S. minnesota and E. coli were tested in a competitive assay with LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support strategies were used to immobilize the protein onto the electrode, one based on electrostatic interactions (electrostatic configuration) and the other trough covalent binding (covalent configuration). LPS from S. minnesota and E. coli were tested in a competitive assay with LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support strategies were used to immobilize the protein onto the electrode, one based on electrostatic interactions (electrostatic configuration) and the other trough covalent binding (covalent configuration). LPS from S. minnesota and E. coli were tested in a competitive assay with LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support proportional to the concentration of the analyte to be detected. We propose a competitive electrochemical assay using a modified gold electrode with a recombinant endotoxin neutralizing protein (ENP) from Saccharomyces cerevisiae as recognition element. Two strategies were used to immobilize the protein onto the electrode, one based on electrostatic interactions (electrostatic configuration) and the other trough covalent binding (covalent configuration). LPS from S. minnesota and E. coli were tested in a competitive assay with LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support strategies were used to immobilize the protein onto the electrode, one based on electrostatic interactions (electrostatic configuration) and the other trough covalent binding (covalent configuration). LPS from S. minnesota and E. coli were tested in a competitive assay with LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support strategies were used to immobilize the protein onto the electrode, one based on electrostatic interactions (electrostatic configuration) and the other trough covalent binding (covalent configuration). LPS from S. minnesota and E. coli were tested in a competitive assay with LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support strategies were used to immobilize the protein onto the electrode, one based on electrostatic interactions (electrostatic configuration) and the other trough covalent binding (covalent configuration). LPS from S. minnesota and E. coli were tested in a competitive assay with LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support LPS labeled with either horseradish peroxidase or biotin. Sensors constructed according to the electrostatic configuration were able to detect the presence of endotoxins in concentrations of 0.2 EU/mL. The results obtained open new routes for a molecular recognition assay for endotoxins. Other possible configurations will be discussed. Fresenius Medical Care Argentina and CONICET are acknowledged for financial support