Patent EP-1845377-A1 (EP)
Inventor: LEHMANN MIRKO DR RER NAT
IPC: G01N33/558
Filed: 2006-04-10
Abstract: The method for determining the concentrations of ligands (9, 10) in a blood sample to be analyzed, comprises binding first and second type of ligand to first receptor (6) and second receptor, and immobilizing receptors on testing sites on a substrate. A third receptor of the second type of receptor is mixed in such a manner with the sample that it is present in a predetermined concentration in the resulting mixture. The components of the mixture, which are not bound to an immobilized receptor, are removed from the substrate. The method for determining the concentrations of ligands (9, 10) in a blood sample to be analyzed, comprises binding first and second type of ligand to first receptor (6) and second receptor, and immobilizing receptors on testing sites on a substrate. A third receptor of the second type of receptor is mixed in such a manner with the sample that it is present in a predetermined concentration in the resulting mixture. The components of the mixture, which are not bound to an immobilized receptor, are removed from the substrate. The mixture is contacted with the substrate in such a manner that first type of ligand can bind to the first receptor and a second type of ligand can bind to the second and third receptors. A first and a second measurement signal that are dependent on the concentration of the ligands bound to the first and the second receptor respectively, are generated. Using the first measurement signal, the concentration of the first type of ligand is determined and with the help of the second measurement signal and the concentration of the third receptor in the mixture, the concentration of the second type of ligand in the sample is determined. For the first and second type of ligand, a specifically-binding detection antibody that is labeled with a first and second marker is contacted with the first and second testing site respectively. The markers that are not bound to an immobilized receptor are removed from the substrate. The first marker and/or the second marker is/are an enzyme, which during the recording of the measurement signals is contacted with two chemicals, between which a chemical redox reaction occurs in the presence of the enzyme. The first measurement signal and/or the second measurement signal is/are generated by measurement of a redox potential. During the recording of the measurement signals, the first marker and/or the second marker are exposed to an activating radiation, which stimulate the marker(s) to give up a luminescent radiation. The first measurement signal and/or the second measurement signal is/are generated by measurement of the luminescence of the concerned marker. During the recording the measurement signals, the first testing site and/or the second testing site is/are contacted with a chemoluminescent substrate. A luminescent radiation is stimulated in dependence to the binding of the first type of ligand to the first receptor and/or to the binding of the second type of ligand to the second receptor. The first measurement signal and/or the second measurement signal is/are generated by measurement of the luminescence at the concerned testing site. Independent claims are included for: (1) a device for determining the concentrations of ligands in a sample to be analyzed; and (2) a kit for the execution of the method.