Procedure And Device For Determining The Concentrations Of At Least Two Ligands

Abstract

In a procedure for determining the concentrations of ligands contained in a sample wherein a first ligand is bond-specific to a first receptor and a second ligand is bond-specific to a second receptor, the first receptor is immobilized on a first test site and the second receptor is immobilized on a second test site on a substrate. A competitor that is bond-specific to the second receptor is mixed with the sample so that it is present in a predetermined concentration in the mixture. The mixture is brought into contact with the substrate so that the first ligand and the second ligand and/or the competitor bind to the first receptor and the second receptor, respectively. Afterwards, the unbound components of the mixture are removed from the substrate. A first metering signal relative to the concentration of the first ligand bound to the first receptor and a second metering signal relative to the concentration of the competitor bound to the second receptor are then generated. The concentrations of the ligands in the sample are determined with reference to the metering signals and the known concentration of the competitor in the mixture.

1 . A procedure for determining the concentrations of at least two ligands presumed to be contained in a sample for analysis, wherein a first ligand is bond-specific for a first receptor and a second ligand is bond-specific for a second receptor, wherein the first receptor is immobilized on at least a first test site and the second receptor is immobilized on at least a second test site on a substrate, wherein a competitor that is bond-specific to the second receptor is mixed with the sample so that said competitor is present in a predetermined concentration in the mixture thus obtained, wherein the mixture is brought into contact with the substrate so that the first ligand can bind to the first receptor and the second ligand and/or the competitor can bind to the second receptor, wherein afterwards components of the mixture that are not bound to an immobilized receptor are removed from the substrate, wherein a first metering signal relative to the concentration of the first ligand bound to the first receptor and a second metering signal relative to the concentration of the competitor bound to the second receptor are then generated, and wherein the concentration of the first ligand in the sample is defined with reference to the first metering signal and the concentration of the second ligand in the sample is determined with reference to the second metering signal and the known concentration of the competitor in the mixture. 
   
   
       2 . A procedure according to  claim 1 , wherein a detection antibody that is bond-specific to the first ligand and marked with a first marker is brought into contact with the first test site, that afterwards markers not bound to an immobilized receptor are removed from the substrate, and that afterwards the first metering signal is generated relative to the concentration of said first marker. 
   
   
       3 . A procedure according to  claim 1 , wherein the competitor is marked with a second marker, and that the second metering signal is generated relative to the concentration of the second marker bound to the second receptor after the components of the mixture not bound to an immobilized receptor are removed from the substrate. 
   
   
       4 . A procedure according to  claim 1 , wherein the first marker and/or the second marker are enzymes/is an enzyme, that the enzyme is brought into contact with at least two chemicals during the capture of the metering signal, between which chemicals a chemical redox reaction takes place in the presence of the enzyme, and that the first metering signal and/or the second metering signal is/are generated by measuring a redox potential. 
   
   
       5 . A procedure according to  claim 1 , wherein the first marker and/or the second marker is/are irradiated with excitation radiation during the capture of the metering signal, said irradiation exciting the marker(s) to emit light, and that the first metering signal and/or the second metering signal is/are generated by measuring the light emitted from the respective marker. 
   
   
       6 . A procedure according to  claim 1 , wherein the first test site and/or the second test site during the capture of the metering signal is/are brought into contact with a chemiluminescent substrate, which is excited to emit light relative to the bond of the first ligand to the first receptor and/or relative to the bond of the competitor to the second receptor, and that the first metering signal and/or the second metering signal is/are generated by measuring the light emitted from the respective test site. 
   
   
       7 - 16 . (canceled)