Fluorescent In-Situ Lipoprotein Detection Gel Electrophoresis
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current electrophoretic technologies for detecting apolipoproteins and lipoprotein particles in biological samples are inefficient, as they require extra-gel sample preparation, pre-electrophoresis protein reduction, and non-specific protein dye staining, limiting the ability to simultaneously identify multiple apolipoproteins and reducing diagnostic specificity.
Innovation Solution
A zonal gel electrophoresis method using fluorescently tagged antibodies that bind specifically to apolipoproteins, allowing for in situ detection of lipoprotein particles within an electrophoretic gel, enabling simultaneous measurement of multiple apolipoproteins and lipoproteins in a single lane without the need for transfer protocols or non-specific staining.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If non-specific protein dye staining is used to detect fixed proteins in gel, then detection can be performed, but specificity is limited in the presence of coincident apolipoproteins
Solution Approach 1:
The patent divides the detection process into separate specific antibody-probe steps for each apolipoprotein type rather than using a single non-specific stain. Each antibody is designed to recognize a specific apolipoprotein antigen, enabling precise segmentation of detection targets and eliminating cross-reactivity issues.
Solution Approach 2:
The patent introduces specific antibodies as intermediary agents between the apolipoprotein antigens and the detection system. These antibodies serve as mediators that provide high-specificity recognition of target apolipoproteins, replacing the non-specific protein dye and enabling both precise detection and multiplexing capability.
2Measurement precision
If extra-gel sample preparation and pre-electrophoresis protein reduction are required, then detection can be performed, but time and complexity increase
Solution Approach 1:
The patent performs preliminary conjugation of fluorescent labels to antibodies before the electrophoresis assay, eliminating the need for post-electrophoresis sample preparation steps. This preliminary preparation of reagents streamlines the overall process by moving complex steps to before the main assay rather than requiring them during or after electrophoresis.
Solution Approach 2:
The patent merges the detection reagents (antibodies with fluorescent labels) directly into the electrophoresis gel matrix, combining multiple functions into a single integrated system. This eliminates separate extra-gel preparation steps and allows detection to occur in-situ within the gel during or after electrophoresis without additional manipulation.
3Measurement precision
If transfer protocols are used for detection, then protein analysis can be performed, but device complexity and operational steps increase
Solution Approach 1:
The patent extracts and eliminates the transfer protocol step entirely from the detection process. By incorporating detection antibodies directly into the gel matrix, the method removes the need to transfer proteins from the gel to a separate membrane, simplifying the overall procedure while maintaining detection precision.
Solution Approach 2:
The gel matrix serves multiple functions simultaneously: it acts as both the electrophoresis medium for protein separation and the detection medium containing the fluorescently-labeled antibodies. This self-service approach eliminates the need for separate transfer membranes and protocols, reducing device complexity while preserving analytical capability.
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 method provides more efficient and specific detection of apolipoproteins and lipoproteins, reducing time, labor, and costs, while enabling the simultaneous analysis of multiple particles, thereby improving diagnostic capabilities for cardiovascular disease risk assessment.
Implementation Method 1
the antibody is bound to a first fluorescent signal producing molecule capable of producing or causing production of a first detectable signal
Implementation Method 2
separating the lipoprotein particles present in the biological sample by depositing the biological sample on an electrophoretic gel and carrying out gel electrophoresis
Data Source
Figure 1~2
Figure 3A~3C
Figure 4A~4D
AI summary
The present invention relates to, among other things, a gel electrophoresis system for detecting the level of specific Apolipoproteins and/or lipoprotein particles present in intact lipid particles in a biological sample. The system includes a gel substrate to receive a biological sample, at least two lipoprotein-binding complexes. Each complex includes an antibody that binds a lipoprotein particle or a portion thereof, which is bound to a signal producing molecule capable of producing or causing production of a detectable signal. The system also includes a device for detecting the detectable signal. The present invention also relates to methods of assessing the level of specific Apolipoproteins and/or lipoprotein particles present in a biological sample, determining whether a subject is at increased risk for cardiovascular disease, and monitoring the risk for developing cardiovascular disease.