Kinetic Parameter Determination via Dissociation Phase Extraction
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Solution Overview
Problem
Existing methods for determining kinetic parameters of reactions between analytes and ligands attached to a flow cell are prone to inaccuracies due to artefacts introduced by refractive index changes during sample fluid flow, which affect the entire binding curve used for analysis.
Innovation Solution
The method focuses on using mainly the dissociation phase of the binding curve, with optional inclusion of a small part of the association phase, to determine kinetic parameters, thereby minimizing the impact of artefacts caused by refractive index changes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If the entire binding curve (including association phases) is used to determine kinetic parameters, then more data is available for analysis, but artefacts from refractive index changes introduce inaccuracies
Solution Approach 1:
The patent extracts and uses only the dissociation phase data from the binding curve, excluding the association phase data that contains refractive index artefacts. This selective extraction eliminates the harmful factors while retaining the useful kinetic information needed for accurate parameter determination.
Solution Approach 2:
The binding curve is segmented into distinct phases (association and dissociation), and the patent selectively applies different analysis approaches to each phase. The dissociation phase is used for kinetic parameter determination while the association phase is excluded, creating a segmented analysis strategy that avoids artefact-contaminated data.
2Measurement precision
If only the dissociation phase is used to determine kinetic parameters, then artefact influence is reduced, but less data is available for analysis
Solution Approach 1:
The patent converts the limitation of having less data into a benefit by demonstrating that the dissociation phase alone provides sufficient and more reliable information for kinetic parameter determination. The exclusion of artefact-contaminated association phase data actually improves the quality of the analysis despite reducing the total data volume.
3Quantity of substance
If association phase data is included in kinetic parameter determination, then more binding information is captured, but refractive index artefacts contaminate the results
Solution Approach 1:
The patent extracts only the clean dissociation phase data from the complete binding curve, separating it from the association phase data that contains refractive index artefacts. This extraction ensures that kinetic parameter determination is based solely on artefact-free data, maintaining both information quality and quantity sufficient for accurate analysis.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach reduces the influence of artefacts on kinetic parameter determination, leading to more accurate results by concentrating on the dissociation phase and a minimal portion of the association phase of the binding curve.
Implementation Method 1
existing sensors which are used to measure the amount of analyte which is bound to the ligands on the test surface and output a binding curve, are sensitive to changes in refractive index of the volumes of sample fluid
Data Source
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AI summary
According to the present invention there is provided a method for determining the kinetic parameters of a reaction between an analyte and ligands which are attached to a test surface of a flow cell, the method comprising the steps of, (a) flowing a first volume of sample fluid (V1), which contains said analyte, over the test surface, between a first point in time (t1) to a second point in time (t2); (b) flowing a first volume of buffer fluid (Vb1), which is without analyte, over the test surface, between a third point in time (t3) to a fourth point in time (t4); (c) flowing at least a second volume of sample fluid (V2), which contains said analyte, over the test surface, between a fifth point in time (t5) to a sixth point in time (t6); (d) flowing at least a second volume of buffer fluid (Vb2), which is without analyte, over the test surface, between a seventh point in time (t7) to a eight point in time (t8); (e) using a sensor to measure the binding of analyte to ligands on the test surface to obtain a binding curve; (f) using the only the parts of the binding curve which are in predefined interval time periods, without using the other parts of the binding curve, to determine the kinetic parameters.