Dual Blocking Agent Kit for Antibody Interference in Biological Assays
Solution Overview
Problem
Current fluorescence detection methods for biological samples, particularly those using surface plasmon excitation, face challenges in achieving high measurement accuracy due to the influence of antibodies like anti-serum albumin antibodies present in blood samples.
Innovation Solution
A kit and method utilizing labeled particles with a first binding substance and a first blocking agent, combined with a substrate having a second binding substance and a second blocking agent, where the blocking agents are different, to enhance the accuracy of quantitative determination by avoiding antibody interference.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a single blocking agent is used in fluorescence detection methods, then the measurement sensitivity is improved, but the measurement accuracy deteriorates due to antibody interference
Solution Approach 1:
The patent divides the blocking agent into two distinct types: a first blocking agent that specifically binds to the substance to be measured, and a second blocking agent that prevents non-specific binding. This segmentation allows each blocking agent to perform its specific function independently, resolving the contradiction between sensitivity and accuracy by eliminating antibody interference through the specialized second blocking agent while maintaining detection capability through the first blocking agent
Solution Approach 2:
The patent introduces a second blocking agent as an intermediary substance that mediates between the antibody in the sample and the detection system. This intermediary prevents harmful antibody-antigen interactions that would cause false positives, thereby improving measurement accuracy without affecting the detection of the target substance by the first blocking agent
2Measurement precision
If surface plasmon excitation is used to enhance fluorescence detection, then the detection sensitivity is improved, but the measurement reproducibility deteriorates
Solution Approach 1:
The patent changes the chemical parameters of the blocking agents by selecting two different types with distinct binding specificities. This parameter change allows the system to maintain the high sensitivity provided by surface plasmon excitation while improving reproducibility through the consistent, specific binding behavior of the two-blockage system that eliminates variable antibody interference
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 allows for high-accuracy quantitative determination of substances in biological samples by minimizing the impact of antibodies, thereby improving measurement reproducibility and sensitivity.
Implementation Method 1
a method utilizing the effect of electric field enhancement by plasmon resonance has been known. In this method, in order to generate plasmon resonance, a sensor chip including a metal film provided in a predetermined region on a transparent support is prepared. The excitation light is allowed to enter an interface between the support and the metal film from a side opposite to the metal film formation surface of the support at an angle equal to or greater than a total reflection angle. Surface plasmons are generated in the metal film by irradiation with the excitation light, fluorescence is enhanced by the electric field enhancement effect due to generation of the surface plasmons
Implementation Method 2
a labeled particle that has a first binding substance capable of binding to the substance to be measured... a substrate that has a detection region having a second binding substance capable of binding to any one of the substance to be measured and the first binding substance
Data Source
AI summary
An object of the present invention is to provide a kit and a method by which a quantitative determination of a substance to be measured in a biological sample can be carried out with high accuracy while avoiding the influence of an antibody such as an anti-serum albumin antibody present in blood. According to the present invention, a kit for quantitatively determining a substance to be measured in a biological sample, the kit including: a labeled particle that has a first binding substance capable of binding to the substance to be measured, and a first blocking agent; and a substrate that has a detection region having a second binding substance capable of binding to any one of the substance to be measured and the first binding substance, and a second blocking agent, in which the first blocking agent and the second blocking agent are different from each other, is provided.