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Publication Number
US-8822231-B2
Publication Date
2014-09-02
Expiration Date
Abstract
A method for the analysis of at least two analytes in a liquid sample, in which a substrate is provided wherein at least two different types of capturing molecules are immobilized on the substrate and wherein each type of capturing molecule has specific affinity for an analyte. The sample is contacted with capturing molecules, wherein for at least one analyte to be analyzed contact is induced between the capturing molecules and a labelled detection molecule with specific affinity for the analyte, and for at least one another analyte to be contact is induced between the capturing molecules and a labelled version of the analyte. A detectable signal is measured from the labelled detection molecule and the labelled analyte on the substrate, wherein the concentration of the labelled analyte is adapted to the concentration of the analyte in the sample.
Core Innovation
The invention relates to an assay method and an analysis device for analysis of at least two analytes in a liquid sample. The device includes a substrate with at least two different types of capturing molecules immobilized on the substrate in separately distinct areas, where each type of capturing molecule has specific affinity for the respective analyte.
For a first analyte expected to be in a first concentration, the method adds at least one type of labeled detection molecule having specific affinity for the first analyte, and induces contact between the first analyte bound to or bound to the capturing molecules and the added labeled detection molecule. Measuring the detectable signal from the labeled detection molecules produces a signal that increases with an increasing concentration of the first analyte.
For a second analyte expected to be in a second concentration much higher than the first concentration, the method adds a labeled version or a labeled fragment of the second analyte and induces contact between the capturing molecules, the second analyte, and the labeled version of the second analyte. Measuring the detectable signal from the labeled second analyte produces a detectable signal that decreases with an increasing concentration of the second analyte, and the concentration of the labeled analyte is adapted to the concentration of the unlabelled analyte in the liquid sample.
Claims Coverage
The provided content contains one independent claim covering a method for simultaneous multi-analyte analysis using distinct immobilized capture-molecule areas and concentration-dependent selection of labeled assay formats, resulting in analyte-specific signal trends.
Distinct immobilized capturing molecule types in separately distinct areas
Providing an analysis device comprising a substrate wherein at least two different types of capturing molecules are immobilized on the substrate in separately distinct areas and wherein each type of capturing molecule has specific affinity for the first and second analytes, respectively.
Sandwich-format labeled detection for a lower-concentration analyte with increasing signal
For the first analyte to be analyzed and expected to have the first concentration: adding at least one type of labeled detection molecule having a specific affinity for the first analyte to be analyzed and inducing contact between the first analyte and the added labelled detection molecule; and measuring a detectable signal from labelled detection molecules bound to the first analyte and capturing molecules wherein the detectable signal increases with an increasing concentration of the first analyte.
Competitive-format labeled second analyte with decreasing signal and adapted labeled analyte concentration
For the second analyte to be analyzed and expected to have the second concentration: adding a labeled version or a labeled fragment of the second analyte and inducing contact between the capturing molecules, the second analyte, and the labelled version of the second analyte; and measuring a detectable signal from the labelled second analyte on the substrate wherein the detectable signal decreases with an increasing concentration of the second analyte to be analyzed, wherein the concentration of the labelled analyte is adapted to the concentration of the unlabelled analyte in the liquid sample.
The independent claim combines distinct immobilized capturing-molecule areas with format selection based on analyte concentration: increasing signal with a labeled detection molecule for the first analyte and decreasing signal with a labeled second analyte or labeled fragment for the second analyte, with the labeled-analyte concentration adapted to the unlabelled analyte concentration.
Stated Advantages
Enables binding to equilibrium for precision [procedural detail omitted for safety].
Avoids sample splitting/dilution.
Avoids transducer gain adjustment.
Avoids added diluent/affinity enhancers.
Documented Applications
Use in microfluidic chips to measure CRP with NTproBNP.
Use in microfluidic chips to measure CRP with cTnI.
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