Multiplex DNA Immuno-Sandwich Assay for Distant Protein Interaction Detection
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for detecting interactions between cells, organelles, and large structures are limited in sensitivity and efficiency, particularly in detecting distant interactions and multiple protein-protein interactions in a high-throughput manner.
Innovation Solution
The method involves tagging antibodies with DNA oligo tags that cannot ligate to each other, using an adapter insert to facilitate ligation and amplification of a DNA product, allowing for the detection and quantification of interactions between analytes through Next Generation Sequencing or other analytical methods.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If current methods are used to detect interactions between cells and large structures, then detection can be performed, but sensitivity and efficiency are limited
Solution Approach 1:
The patent introduces an adapter insert as an intermediary DNA molecule that mediates between antibody oligo tags and the amplification system. This adapter enables efficient signal transduction from the antibody-antigen interaction to the DNA amplification readout, significantly enhancing both sensitivity and efficiency of detection
Solution Approach 2:
The patent replaces traditional mechanical/direct detection methods with a molecular biology-based amplification system. By converting the antibody interaction signal into a DNA amplification signal, the system achieves exponential signal amplification, dramatically improving detection sensitivity and throughput
2Length of stationary object
If traditional assays are used, then protein interactions can be detected, but they cannot detect distant interactions or large protein complexes
Solution Approach 1:
The patent extends the detection capability into the spatial dimension by using an adapter insert of variable length. This allows the system to bridge distant antibody binding sites on large protein complexes or cells, enabling detection of interactions that span nanometer to micrometer distances while maintaining detection accuracy through specific molecular recognition
3Adaptability or versatility
If single-plex assays are used, then specific protein interactions can be detected, but multiple interactions cannot be detected simultaneously
Solution Approach 1:
The patent creates a universal detection platform where different antibody pairs can be used with the same adapter insert and amplification system. The adapter serves multiple functions: bridging antibodies, enabling amplification, and allowing multiplexing. This universal design enables simultaneous detection of multiple protein interactions without proportionally increasing system complexity
Solution Approach 2:
The patent uses parameter changes in the form of different adapter inserts with varying lengths or sequences to enable multiplexing. By changing adapter parameters rather than the entire assay system, multiple interactions can be detected simultaneously while keeping the core methodology simple and reusable
4Productivity
If manual assays are used, then detection can be performed, but high-throughput automation is difficult
Solution Approach 1:
The patent employs self-service principles through the self-ligating capability of the adapter insert and antibody oligo tags. The molecular components automatically assemble through complementary base pairing and ligation without requiring manual intervention, enabling straightforward automation of the assay process for high-throughput applications
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 significantly enhances sensitivity and efficiency, enabling the detection of multiple protein-protein interactions and protein modifications in a multiplex format, with the ability to analyze distant interactions and large protein complexes, and can be performed in a high-throughput 96-well format.
Implementation Method 1
adding a ligase to the mixture from step (b), resulting in a ligated single DNA Product comprised of the double-stranded DNA oligo tags, and the double-stranded adapter-insert DNA
Implementation Method 2
amplifying the ligated single DNA product, or portions thereof
Data Source
Figure 1
Figure 2
Figure 3
AI summary
This invention is directed to a method for detecting near and distant interactions between cells, organelles and other large structures. This method can also be used for specific detection and quantification of protein-protein and macro-molecular interactions, as well as quantitative analysis of multiple proteins and their modifications.