Artificial Transmembrane Protein for Label-Free Biomolecular Detection
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Solution Overview
Problem
Current methods for detecting intracellular or intravesicular biomolecular interactions, such as those involving G-protein coupled receptors, are limited by the need for fluorescent labeling, which can alter cellular physiology and provide incomplete or cross-sensitive data, especially in complex environments like living cells.
Innovation Solution
An artificial transmembrane protein with an extracellular binder structure, a hydrophobic transmembrane domain, and an intracellular or intravesicular receptor structure, designed to interact specifically with intracellular or intravesicular components, allowing for label-free detection using an evanescent illuminator with a template nanopattern to generate a transmembrane nanopattern and reduce cross-sensitivity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If fluorescent labeling is used to detect intracellular biomolecular interactions, then detection sensitivity is improved, but cellular physiology is altered and measurement accuracy deteriorates
Solution Approach 1:
The invention extracts and removes the fluorescent label from the detection system, replacing it with label-free detection methods. The transmembrane protein structure enables direct detection of biomolecular interactions without requiring fluorescent tags, thereby eliminating the distortion caused by labeling while maintaining detection capability
Solution Approach 2:
The transmembrane protein acts as an intermediary structure that spans the cell membrane, with its intracellular domain detecting target molecules and its extracellular domain enabling optical detection. This mediator allows indirect detection of intracellular interactions through the cell membrane without penetrating or altering cellular physiology
2Difficulty of detecting and measuring
If fluorescent labels are attached to target samples, then binding detection capability is improved, but additional preparation steps and costs increase
Solution Approach 1:
The invention extracts the fluorescent label requirement entirely from the detection process. The transmembrane protein's intrinsic optical properties and the evanescent field interaction enable direct detection of binding events without any labeling steps, eliminating both preparation complexity and associated costs
Solution Approach 2:
The transmembrane protein structure itself provides the detection capability through its natural interaction with evanescent light fields. The system is self-sufficient, requiring no external fluorescent tags or labeling reagents, as the protein structure enables direct optical detection of biomolecular interactions
3Adaptability or versatility
If conventional detection methods are used in complex cellular environments, then general applicability is maintained, but cross-sensitivity and false signals increase
Solution Approach 1:
The invention creates a localized detection environment using the evanescent field that penetrates only a few hundred nanometers into the cellular environment. This localized probing region restricts detection to specific molecular interactions at the transmembrane protein location, eliminating cross-sensitivity from distant cellular components while maintaining general applicability
Solution Approach 2:
The transmembrane protein segments the detection function into distinct domains: the intracellular domain for specific target recognition and the extracellular domain for optical interaction. This segmentation allows specific binding detection in complex environments by separating the recognition function from the detection function, reducing false signals from non-specific interactions
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 direct and specific monitoring of biomolecular interactions with reduced cross-sensitivity, allowing for real-time measurement of molecular mass and number of receptors involved, providing a more accurate and non-invasive assessment of intracellular processes.
Implementation Method 1
an artificial transmembrane protein for use in a biomolecular detection device for detecting intracellular or intravesicular biomolecular interactions
Implementation Method 2
a hydrophobic transmembrane domain
Data Source
AI summary
Disclosed herein is an artificial transmembrane protein for use in a biomolecular detection device for detecting intracellular or intravesicular biomolecular interactions, the artificial transmembrane protein having an extracellular or extravesicular binder structure, a hydrophobic transmembrane domain, and an intracellular or intravesicular domain with an intracellular or intravesicular receptor structure, wherein the receptor structure is configured to interact with an intracellular or intravesicular component of the biomolecular interaction to be detected and wherein the extracellular or extravesicular binder structure is configured to bind to membrane recognition elements arranged along a plurality of predetermined lines of the biomolecular detection device.