Patent EP-2264461-A1 (EP)
Inventor: MIETHE PETER
IPC: G01N30/60
Filed: 2009-06-19
Abstract: The device (10) comprises a sample analysis vessel (12), which has an upper end with a sample inlet opening (14) and a lower end with a sample outlet opening (16), where the sample analysis vessel is flowed-through by an aqueous sample to be analyzed from the sample inlet opening up to the sample outlet opening. The sample analysis vessel is provided with a sample chamber subsequent to the sample inlet opening for receiving the sample and a marking agent carrier (20) that is arranged downstream to the sample chamber and has a porous solid phase and marking agent in a dried state. The device (10) comprises a sample analysis vessel (12), which has an upper end with a sample inlet opening (14) and a lower end with a sample outlet opening (16), where the sample analysis vessel is flowed-through by an aqueous sample to be analyzed from the sample inlet opening up to the sample outlet opening. The sample analysis vessel is provided with a sample chamber subsequent to the sample inlet opening for receiving the sample, a marking agent carrier (20) that is arranged downstream to the sample chamber and has a porous solid phase and marking agent in a dried state and an affinity ligand carriers that are arranged downstream to the marking agent carrier and have a porous solid phase with immobilized affinity ligands and optionally further pre-reaction reactants. The flow resistance of the marking agent carrier is in ratio to the flow resistance of the arrangement present downstream to the marking agent carrier, so that first partial volumes of the sample rapidly flow through the marking agent carrier than a porous flow-through element present downstream to the marking agent carrier, which is adjacent to the flow-through element, where the flow-through element is affinity ligand carrier or a spacer element for spacing the affinity ligand carrier from the marking agent carrier. The flow resistance of the marking agent carrier is less than the flow resistance of the flow-through element present downstream to the marking agent carrier, where the flow resistances of the marking agent carrier and the flow-through element are defined by their permeability coefficients. The permeability coefficient of the marking agent carrier is greater than the permeability coefficient of the flow-through element, where the permeability coefficient of the marking agent carrier amounts to 1.5-2.5 times of the permeability coefficient of the flow-through element. The marking agent carrier is formed from the solid phase having fibers, which are aligned in longitudinal extension. The solid phase has a pore portion of 20-80% with an average pore diameter of 5-200 mu m. The affinity ligand carrier has a sintered substrate made of polyolefin or glass. The total volume of the sample is equal to the sum of the first partial volume and second partial volume, which initially remains in the sample chamber after filling the marking agent carrier with the first volume of the sample. The second partial volume is equal to 110-120% of total pore volumes of all of the elements of the sample vessel to be flowed through by the sample for the purpose of the affinity-chromatographic sample test. The first partial volume of the sample reacts with the marking agent during flow through the sample analysis vessel and optionally reacts with further pre-reaction reactants or mixes with the marking agent in a reproducible manner. The vessel is provided with several porous spacer elements for spacing the adjacent affinity ligand carrier. The flow resistance of the spacer element is different from the flow resistance of the affinity ligand carrier. Three affinity ligand carriers are provided with four spacer elements arranged alternatively to the affinity ligand carriers. The flow resistances of the spacer elements and the affinity ligand carriers are defined by their permeability coefficients. The permeability coefficient of the spacer elements is less or greater than the permeability coefficient of the affinity ligand carriers, where the permeability coefficient of the affinity ligand carriers amounts to 1.5-0.75 times of the permeability coefficient of the spacer elements. The spacer elements consist of sintering material. A refuse container is provided and is arranged below the outlet opening of the sample analysis vessel or the outlet opening of the sample analysis vessel is provided with a closure element automatically closing during wetting with the liquid. The affinity ligands consist of substances including proteins such as antibodies, protein A, protein G and streptavidin, native or recombinant surface antigens of microorganisms, somatic cells, viruses, lectins, nucleic acids such as single-stranded RNA and DNA probes, aptamers, oligomeric compounds such as peptides, oligosaccharides or low molecular compounds such as haptens and sugars. The marking agent is produced by conjugation of proteins, nucleic acids, oligopeptids or haptens with a marking molecule, which is detectable optically such as photometrically, fluorometrically, luminometrically or phosphorimetrically, or magnetically or electrochemically such as amperometrically, potentiometrically and conductometrically. The flow resistance and/or the permeability coefficient of the marking agent carrier, the affinity ligand carrier and the spacer element are predetermined by selecting the pore size and/or the wetting properties or interfacial tension i.e. the interaction energy of the surface of the solid phase to the liquid phase.