Label-Free Protein Interaction Detection via Diffusion in Microscopic Reservoirs
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Solution Overview
Problem
Current methods for detecting and characterizing protein interactions are limited by the need for labeling, tethering, and are not suitable for cycling measurements, which can alter protein functionality and are not sensitive enough to detect proteins at undetectable concentrations.
Innovation Solution
The use of a combination of spheroids, magnetic fields, fluorescence, optics, filter technology, gel technology, electrochemistry, and Matrix-Assisted Laser Desorption Ionization (MALDI) to detect and characterize analyte particles by measuring their diffusion coefficients and interactions within microscopic reservoirs, allowing label-free and tether-free analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If labeling and tethering methods are used to detect protein interactions, then detection sensitivity can be improved, but protein functionality is altered and measurements cannot be cycled
Solution Approach 1:
The invention extracts the detection function from the protein itself by using a separate indicator species that reports on the presence and interaction of target proteins through diffusion-based measurement, eliminating the need for labeling or tethering the proteins of interest
Solution Approach 2:
The invention introduces an intermediary indicator species that mediates the detection process by undergoing diffusion-based measurement to report on protein interactions, allowing indirect detection without direct modification of the target proteins
2Difficulty of detecting and measuring
If labeling methods are used to detect proteins, then detection capability is enhanced, but the labeling process interferes with protein interactions and solubility
Solution Approach 1:
The invention extracts the detection function from the protein by using a separate indicator species that reports on protein presence and interactions through diffusion-based measurement, eliminating harmful labeling effects
Solution Approach 2:
The invention replaces chemical labeling methods with a physical diffusion-based measurement system, substituting chemical modification with a physical process that does not interfere with protein structure or function
3Device complexity
If conventional diffusion measurements are used, then measurement simplicity is maintained, but sensitivity is insufficient for undetectable protein concentrations
Solution Approach 1:
The invention implements nested measurement by having the indicator species nested within or associated with the target protein complex, allowing the diffusion measurement of the larger complex to report on the presence and interaction of the target protein at very low concentrations
Solution Approach 2:
The invention creates a measurable copy of the interaction event by using an indicator species that replicates the diffusion behavior of the target protein complex, allowing indirect measurement of proteins at undetectable concentrations
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 enables sensitive, label-free, and tether-free detection of protein interactions, allowing for repeated measurements and analysis of proteins in their native state, even at low concentrations, and is applicable to complex aqueous mixtures and opaque samples.
Implementation Method 1
the means of trapping is terminated and the analyte particles diffuse out of the reservoirs
Implementation Method 2
applying an electric field through said through holes containing the analyte particles; measuring a change in electric current flow with time indicative of the diffusion rates of said analyte particles
Implementation Method 3
Matrix-Assisted Laser Desorption Ionization (MALDI) to detect and characterize analyte particles
Data Source
AI summary
Methods for the detection of biologically relevant molecules that comprise concentrating such molecules into microscopic holes in a sheet of chemically inert material, restricting the openings, and measuring the electric current through the holes or the fluorescence near the hole openings. The electric current or fluorescence will change as the molecules diffuse out of the holes, providing a measure of the diffusion rate and thereby detecting the presence and characteristics of the molecules. For molecules that interact, the diffusion rate will be slower than for molecules that do not interact, yielding a determination of the molecular interaction. Capping the population of holes and inserting into a mass spectrometer allows identification of the molecules.