Oxidized LDL Evaluation Apparatus Kinetic Assay
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Solution Overview
Problem
Current methods for measuring oxidized LDL, such as the sandwich ELISA method, are complex and cannot perform rapid measurements.
Innovation Solution
An evaluation apparatus and method that acquire information on the change amount of absorbance per unit time by causing a peroxidase to act on hydrogen peroxide produced by solubilizing LDL in a specimen with a surfactant, and subjecting the resultant to an oxidation reaction with cholesterol oxidase, to introduce hydrogen peroxide to a color reaction, followed by measurement of the change of absorbance with time.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If the sandwich ELISA method is used to measure oxidized LDL, then measurement precision is improved, but device complexity and measurement time increase
Solution Approach 1:
The patent extracts the essential detection function from the complex sandwich ELISA method by using a simplified kinetic assay that measures only the rate of color formation rather than requiring multiple washing steps, blocking procedures, and complex reagent combinations. This extraction maintains measurement precision while dramatically reducing procedural complexity.
Solution Approach 2:
The patent replaces the mechanical complexity of the ELISA system (requiring plate-based manual operations, multiple reagent additions, and complex incubation protocols) with an automated kinetic measurement system that uses a single reagent addition followed by continuous absorbance monitoring, substituting mechanical complexity with automated optical measurement.
2Measurement precision
If the sandwich ELISA method is used to measure oxidized LDL, then measurement precision is improved, but measurement time increases
Solution Approach 1:
The patent applies preliminary action by pre-mixing all necessary reagents into a single kinetic assay solution that contains everything needed for the measurement. This eliminates the need for multiple sequential reagent additions and incubation steps required in ELISA, allowing the measurement to start immediately after a single addition and complete rapidly through continuous kinetic monitoring.
Solution Approach 2:
The patent implements continuous measurement of absorbance change over time during the kinetic reaction, eliminating the discontinuous stepwise protocol of ELISA. The useful action of measuring oxidized LDL continues uninterrupted through automated kinetic monitoring, allowing rapid data collection without the time-consuming intermediate steps of ELISA washing and development.
3Productivity
If a simple measurement method is used, then measurement speed is improved, but measurement precision deteriorates
Solution Approach 1:
The patent changes the measurement parameter from endpoint absorbance (ELISA) to kinetic rate of absorbance change. By measuring the slope of absorbance increase over time rather than the final color intensity, the method achieves both speed and precision: the kinetic rate provides a more sensitive and specific indicator of oxidized LDL presence while enabling rapid automated measurement.
Solution Approach 2:
The patent incorporates continuous feedback through automated monitoring of absorbance change over time. The system continuously measures the kinetic rate and uses this feedback to determine oxidized LDL concentration, allowing real-time adjustment and ensuring accurate results. This feedback mechanism maintains precision while enabling high-speed automated measurement.
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
Enables rapid evaluation of the amount of oxidized LDL in a specimen based on the change amount of absorbance per unit time, providing a more efficient and accurate indicator for diseases related to vascular endothelial cell function.
Implementation Method 1
solubilizing an LDL in a specimen with a surfactant that acts on the LDL
Implementation Method 2
subjecting the resultant to an oxidation reaction with a cholesterol oxidase, to introduce the hydrogen peroxide
Implementation Method 3
causing a peroxidase to act on hydrogen peroxide
Implementation Method 4
introduce the hydrogen peroxide to a color reaction, followed by measurement of a change of the absorbance with time
Data Source
AI summary
An evaluation apparatus includes an acquiring unit configured to acquire information on a change amount of an absorbance per unit time. The change amount is obtained by a measurement method including causing a peroxidase to act on hydrogen peroxide, which is produced by solubilizing a low-density lipoprotein (LDL) in a specimen with a surfactant that acts on the LDL, and subjecting the resultant to an oxidation reaction with a cholesterol oxidase, to introduce the hydrogen peroxide to a color reaction, followed by measurement of a change of the absorbance with time. The evaluation apparatus also includes an evaluating unit configured to evaluate an amount of an oxidized LDL in the specimen based on the change amount.