Labeled Complex for Immunoassay Sensitivity
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Solution Overview
Problem
Existing methods for labeling antigens in immunodetection often result in reduced immunological activity and instability of labeled complexes, leading to false positives and low sensitivity, particularly due to direct labeling techniques and the complexity of indirect labeling methods that require specific micromolecules or marker proteins.
Innovation Solution
A labeled complex is created by coupling a marker protein with functional groups to an antigen, followed by secondary labeling with a signal generation substance, using methods like covalent coupling reactions to maintain antigen conformation and enhance signal amplification, thereby improving sensitivity and stability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If direct labeling technique is used to label antigen with marker, then labeling process is simple, but immunological activity of antigen is reduced and labeled complex is unstable
Solution Approach 1:
The labeling process is divided into two independent stages: first coupling the marker protein to the antigen to form a labeled-complex intermediate, then separately coupling the signal generation substance to the marker protein. This segmentation allows each coupling reaction to be optimized independently, maintaining antigen stability while achieving effective labeling.
Solution Approach 2:
The marker protein serves as an intermediary between the antigen and the signal generation substance. By introducing this intermediate component, the direct interaction between antigen and signal molecule is avoided, preventing damage to antigen immunological activity while still enabling signal detection.
2Reliability
If indirect labeling method with micromolecule or marker protein is used, then immunological activity is maintained, but reagent composition becomes complex
Solution Approach 1:
The marker protein performs multiple functions simultaneously: it serves as a coupling bridge between antigen and signal molecule, provides multiple coupling sites for signal generation substances, and maintains antigen conformation. This multi-functionality reduces the need for separate components required in traditional indirect labeling methods.
Solution Approach 2:
The invention creates a composite labeled complex structure where marker protein, antigen, and signal generation substance are covalently combined. This composite structure integrates the benefits of indirect labeling (maintained immunological activity) with simplified composition by making the marker protein the central organizing element.
3Measurement precision
If multiple signal generation substances are coupled to marker protein, then detection sensitivity is enhanced through signal amplification, but labeling process becomes more complex
Solution Approach 1:
The marker protein is first coupled to the antigen to create a stable labeled-complex intermediate with multiple available coupling sites. This preliminary action prepares the structure to receive multiple signal generation substances in subsequent steps, simplifying the overall process by establishing the framework first.
Solution Approach 2:
The labeling process uses a nested approach where the marker protein is first attached to the antigen, then multiple signal generation substances are attached to the marker protein. This creates a nested structure (signal substance within marker protein within antigen complex) that efficiently organizes multiple components.
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 maintains antigen immunoreactivity, enhances detection sensitivity, and stabilizes the labeled complex, addressing the limitations of direct and indirect labeling methods while simplifying the reagent composition.
Implementation Method 1
coupling a marker protein with functional groups to an antigen, followed by secondary labeling with a signal generation substance, using methods like covalent coupling reactions
Data Source
Figure 1
AI summary
A labeled complex, the preparation method thereof, a kit containing the same, the application of the kit and a detection system comprising the kit are provided. The labeled complex comprises: an antigen; a marker protein coupled with the antigen to form a labeled-complex intermediate; and a signal generation substance coupled with the labeled-complex intermediate to form the labeled complex. By applying the technical solutions provided herein to couple a marker protein with a functional group of an antigen, the conformation of the antigen is maintained, or changed without covering an epitope, thus keeping the immune reactivity of the antigen. Moreover, by labeling an antigen with marker proteins, one antigen can carry a plurality of marker proteins, each of which is bound with a plurality of signal generation substances during a secondary signal generation substance labeling process to generate an effective signal amplification effect, thus further improving the detection sensitivity.