Diagnostic methods
First Claim
1. A non-invasive method for the detection of the presence or absence of a Helicobacter pylori antigen in a biological sample comprising:
- a) contacting a biological sample obtained from a patient suffering or suspected of suffering from H. pylori infection with a biosensor comprising (i) a carrier substrate which has a solid surface, (ii) antibodies against H. pylori, or antigen-binding fragments-thereof, and (iii) hydrophilic biomolecule-repellent molecules;
wherein;
the solid surface of the carrier substrate produces a surface plasmon resonance (SPR) signal;
both the antibodies or antigen-binding fragments thereof and the hydrophilic biomolecule-repellent molecules comprise a functional group having affinity for the solid surface of the carrier substrate;
both the antibodies or antigen-binding fragments thereof and the hydrophilic biomolecule-repellent molecules are covalently attached directly to the solid surface of the carrier substrate through the functional group having affinity for the solid surface of the carrier substrate, thereby forming a monolayer;
both the antibodies or antigen-binding fragments thereof and the hydrophilic biomolecule-repellent molecules are self-assembled on the solid surface of the carrier substrate; and
the antibodies or antigen binding fragments thereof are attached to the solid surface of the carrier substrate at the same time as, or prior to, attachment of the hydrophilic biomolecule-repellent molecules to the solid surface of the carrier substrate; and
b) detecting the signal resulting from the formation of an antibody-antigen-complex;
wherein the hydrophilic biomolecule-repellent molecules are neutral hydrophilic monomers or polymers selected from the group consisting of polyacrylamide, poly-N,N-dimethylacrylamide, and N-[tris(hydroxymethyl)methyl]acrylamide.
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Accused Products
Abstract
The present invention relates to novel methods for the diagnosis of Helicobacter pylori infection. Specifically, the present invention relates to novel non-invasive methods for the detection of the presence or absence of a Helicobacter pylori antigen or a metabolite produced by the bacterium in a biological sample with a biosensor-based measurement. The present invention also related to the use of a biosensor containing specific antibodies against H. pylori or antigen-binding fragments thereof immobilized thereto together with biomolecule-repellent polymers preventing the non-specific binding. The invention also relates to test kits useful in the methods.
60 Citations
16 Claims
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1. A non-invasive method for the detection of the presence or absence of a Helicobacter pylori antigen in a biological sample comprising:
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a) contacting a biological sample obtained from a patient suffering or suspected of suffering from H. pylori infection with a biosensor comprising (i) a carrier substrate which has a solid surface, (ii) antibodies against H. pylori, or antigen-binding fragments-thereof, and (iii) hydrophilic biomolecule-repellent molecules;
wherein;the solid surface of the carrier substrate produces a surface plasmon resonance (SPR) signal; both the antibodies or antigen-binding fragments thereof and the hydrophilic biomolecule-repellent molecules comprise a functional group having affinity for the solid surface of the carrier substrate; both the antibodies or antigen-binding fragments thereof and the hydrophilic biomolecule-repellent molecules are covalently attached directly to the solid surface of the carrier substrate through the functional group having affinity for the solid surface of the carrier substrate, thereby forming a monolayer; both the antibodies or antigen-binding fragments thereof and the hydrophilic biomolecule-repellent molecules are self-assembled on the solid surface of the carrier substrate; and the antibodies or antigen binding fragments thereof are attached to the solid surface of the carrier substrate at the same time as, or prior to, attachment of the hydrophilic biomolecule-repellent molecules to the solid surface of the carrier substrate; and b) detecting the signal resulting from the formation of an antibody-antigen-complex; wherein the hydrophilic biomolecule-repellent molecules are neutral hydrophilic monomers or polymers selected from the group consisting of polyacrylamide, poly-N,N-dimethylacrylamide, and N-[tris(hydroxymethyl)methyl]acrylamide. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11)
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12. A test kit for the detection of the presence or absence of a H. pylori antigen in a biological sample, the test kit comprising:
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a) a biosensor comprising;
(i) a carrier substrate which has a solid surface, (ii) antibodies against H. pylori or antigen-binding fragments thereof, and (iii) hydrophilic biomolecule-repellent molecules;
wherein;the solid surface of the carrier substrate produces a surface plasmon resonance (SPR) signal; both the antibodies or antigen-binding fragments thereof and the hydrophilic biomolecule-repellent molecules comprise a functional group having affinity for the solid surface of the carrier substrate; both the antibodies or antigen-binding fragments thereof and the hydrophilic biomolecule-repellent molecules are covalently attached directly to the solid surface of the carrier substrate through the functional group having affinity for the solid surface of the carrier substrate, thereby forming a monolayer; and both the antibodies or antigen-binding fragments thereof and the hydrophilic biomolecule-repellent molecules are self-assembled on the solid surface of the carrier substrate; and the antibodies or antigen binding fragments thereof are attached to the solid surface of the carrier substrate at the same time as, or prior to, attachment of the hydrophilic biomolecule-repellent molecules to the solid surface of the carrier substrate, and b) reagents needed for the calibration and quality control of the detection; and c) auxiliary reagents; wherein the hydrophilic biomolecule-repellent molecules are neutral hydrophilic monomers or polymers selected from the group consisting of polyacrylamide, poly-N,N-dimethylacrylamide, and N-[tris(hydroxymethyl)methyl] acrylamide molecules. - View Dependent Claims (13, 14, 15, 16)
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Specification