Cell Detection via Reporter Gene Fluorescence
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for selecting cells producing a polypeptide of interest, such as hybridoma clones or transfected cells, are time-consuming and rely on visual colony size, failing to differentiate between high and low-producing cells, and require lengthy growth phases for amplifiable selectable markers.
Innovation Solution
A method involving a marker compound that associates with a reference polypeptide, allowing for rapid identification of cells producing a polypeptide of interest through optical imaging, without the need for specific antibodies, by detecting the association of the marker compound with cells, which can be amplified in the presence of the marker.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If robot implemented cloning method is used to pick individual colonies, then picking speed increases to up to 400 clones per hour, but the method cannot discriminate between high-producing and low-producing hybridoma cells since it relies solely on visualisation of colony size
Solution Approach 1:
The patent employs fluorescence emission (a form of light/color change) from reporter genes to visually distinguish high-producing cells from low-producing cells. The imaging head captures fluorescence signals that correlate with polypeptide production levels, enabling the robot to select colonies based on productivity rather than just size.
Solution Approach 2:
The patent introduces reporter genes (intermediary elements) that produce detectable signals (fluorescence or colorimetric responses) as proxies for polypeptide production. These reporter genes serve as mediators that translate internal cellular productivity into externally observable signals, allowing indirect measurement of cell productivity.
2Measurement precision
If specific reagents such as antibodies are used to detect polypeptide production, then productivity discrimination is enabled, but the requirement for different reagents for different polypeptides increases device complexity and reagent requirements
Solution Approach 1:
The patent employs universal reporter genes (such as GFP, luciferase, or other broadly applicable reporter systems) that can detect polypeptide production across different target proteins without requiring specific antibodies for each polypeptide. The same imaging system and reporter gene approach works for detecting production of any polypeptide of interest, eliminating the need for polypeptide-specific detection reagents.
3Reliability
If amplifiable selectable markers are used to identify cell clones, then high level of gene product expression can be achieved, but lengthy growth phases are required for adequate distinction between transfected and non-transfected cells
Solution Approach 1:
The patent performs preliminary detection of polypeptide production using reporter genes before committing to lengthy amplification and selection processes. By detecting productivity early through fluorescent or colorimetric signals, the system can identify high-producing clones without requiring extended growth phases for amplifiable marker selection, thus saving time while maintaining reliability.
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 rapid and efficient selection of high-producing cells by linking the production of the polypeptide of interest to the reference polypeptide, reducing the selection time and eliminating the need for lengthy growth phases, using readily available reagents and automated imaging systems.
Implementation Method 1
exposing one or more cells to a marker compound which associates with a reference polypeptide
Implementation Method 2
detected by an automated imaging system
Data Source
AI summary
In one aspect the present invention provides a method of identifying a cell or cell colony which produces a polypeptide of interest, the method comprising exposing one or more cells to a marker compound which associates with a reference polypeptide, wherein production of the polypeptide of interest by the cells is linked to production of the reference polypeptide, and detecting association of the marker compound with the one or more cells, thereby identifying a cell or cell colony which produces the polypeptide of interest.