Biomolecular Interaction Detection via Translocation Module
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Solution Overview
Problem
Current methods for detecting biomolecular interactions, such as yeast two-hybrid, FRET, and Bi-FC, face challenges including difficulty in investigating membrane or nuclear proteins, high probability of false positives, and low success rates due to complex protein positioning requirements.
Innovation Solution
A novel method involving constructs with a bait and a prey, each labeled with a translocation module and a labeling material, allowing real-time detection of interactions by tracing their movement within a cell in response to external or internal signals.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If yeast two-hybrid method is used to detect protein interactions, then large-scale screening is possible, but false positives increase and membrane/nuclear proteins cannot be investigated
Solution Approach 1:
The patent replaces the biochemical yeast two-hybrid system with a fluorescence-based optical detection system. The TMD-EGFP construct uses translocation to the plasma membrane as a readout, substituting the mechanical/biochemical yeast colony color change with optical fluorescence detection, thereby eliminating false positives while maintaining screening capacity
Solution Approach 2:
The patent introduces an intermediary translocation module (TMD) that mediates the interaction detection. The TMD acts as a bridge between the protein interaction event and the detectable signal (fluorescence localization), allowing specific detection of membrane-associated interactions while excluding nuclear proteins that cannot translocate to the plasma membrane
2Measurement precision
If FRET or Bi-FC methods are used to detect protein interactions, then accuracy is improved, but experimental success rate decreases due to complex protein positioning requirements
Solution Approach 1:
The patent employs self-service through the autonomous translocation property of the TMD. The translocation module automatically directs the construct to the plasma membrane in response to external stimuli without requiring manual positioning or complex experimental setup, thereby maintaining high accuracy while simplifying the experimental process and increasing success rates
Solution Approach 2:
The patent introduces dynamic translocation as the detection mechanism. Instead of relying on static protein positioning or complex spatial arrangements required by FRET/Bi-FC, the system uses dynamic translocation to the plasma membrane that can be easily triggered and observed, greatly simplifying experimental implementation while maintaining detection accuracy
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 accurate, real-time monitoring of biomolecular interactions with reduced false positives and increased experimental accuracy, applicable to various cell types and capable of screening for regulators of protein-protein interactions.
Implementation Method 1
a first construct comprising a bait, a first labeling material and a translocation module; (b) detecting the distribution of the first construct and the second construct in the cell
Data Source
AI summary
Disclosed is a novel method for detecting interactions of biomolecules. More particularly, the disclosed method includes (a) preparing a cell comprising (i) a first construct comprising a bait, a first labeling material and a translocation module; and (ii) a second construct comprising a prey and a second labeling material; (b) detecting the distribution of the first construct and the second construct in the cell. the present invention provides a method capable of detecting bindings and interactions occurring in a living cell in real time, and a method for screening a material that alters the binding and the interaction. The method of the present invention overcomes the disadvantages including inaccuracy and complexity of existing biomaterial interaction detection techniques. By labeling both constructs to promote accuracy, the present invention provides a novel real-time, antibody-free analysis.