Microwell Vesicle Screening for Sensitive Target-Cell Detection
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Solution Overview
Problem
Existing methods for screening cells that produce specific binding substances, such as antibodies, suffer from low sensitivity and efficiency, particularly when dealing with large cell populations, and often result in false positives and reduced analysis capacity due to cell damage and well size limitations.
Innovation Solution
A screening method involving immobilizing vesicles with target substances on microwells, introducing candidate cells and labeled substances, and identifying target cells by detecting bound labels, utilizing specific labels and vesicles like Tim family proteins and antibodies for high sensitivity and efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional screening methods are used with large cell populations, then the ability to search for specific binding substances is maintained, but screening sensitivity decreases and false positives increase
Solution Approach 1:
The invention divides the cell population into individual cells within separate microwells, allowing each cell to be independently evaluated. This segmentation enables precise detection of specific binding substances produced by individual cells, thereby improving screening sensitivity and reducing false positives by eliminating background interference from other cells.
Solution Approach 2:
The invention introduces a labeled substance as an intermediary that binds to the objective substance produced by target cells. This labeled substance serves as a detectable mediator, allowing indirect detection of the objective substance through fluorescence or other labels, thereby enhancing measurement precision and reliability.
2Productivity
If conventional microwell sizes are used, then the analysis capacity is limited, but cell damage occurs and screening efficiency decreases
Solution Approach 1:
The invention changes the physical parameters of the microwells by reducing their size to accommodate individual cells. This parameter change allows for higher density of wells per plate, increasing analysis capacity while the smaller well size reduces the overall volume of reagents and media, thereby reducing cell damage from resource depletion and improving screening efficiency.
3Measurement precision
If all or part of protein is immobilized in wells for ELISA, then binding evaluation is performed, but the three-dimensional structure information of cell membrane proteins is not fully utilized
Solution Approach 1:
The invention uses cells themselves as three-dimensional models that naturally display the target protein in its native conformation. Instead of immobilizing denatured or simplified protein fragments, the living cells preserve the complete three-dimensional structure information, allowing simultaneous binding evaluation and structural information retention.
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
The method achieves high sensitivity and efficiency in identifying target cells with reduced false positives and increased analysis capacity by using smaller microwells and specific labeling techniques.
Implementation Method 1
a labeled substance which is a substance that binds to the objective substance
Data Source
AI summary
An object of the present invention is to provide a screening method for a cell, which has excellent screening sensitivity and excellent screening efficiency, a manufacturing method for a nucleic acid, in which the nucleic acid is acquired from a cell obtained by the screening method, and a manufacturing method for an objective substance, in which an objective substance is manufactured using the nucleic acid. According to the present invention, provided is a screening method for a target cell which is a cell that produces an objective substance, the screening method including the following (1) to (3), (1) immobilizing a vesicle having a target substance on a membrane surface in a microwell, (2) introducing a candidate cell obtained from a cell population containing the target cell and a labeled substance which is a substance that binds to the objective substance into the microwell in which the vesicle has been immobilized, and (3) identifying the target cell by detection of a label of the objective substance bound to the target substance.