Cell Support Residue Measurement by Fluorescent Saccharide Detection
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Solution Overview
Problem
Existing methods for large-scale cell culture using cell supports, such as microcarriers, often result in residues from the cell support being collected with the cells, complicating the separation process and making it difficult to measure cell support-derived components accurately.
Innovation Solution
A method for measuring cell support-derived components in a liquid sample using a mixture of the sample, an acid, and an aromatic compound that reacts with saccharides under heating, followed by fluorescence or absorption analysis, allowing for the detection and quantification of polysaccharide cell support residues.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If cell supports are used for large-scale cell culture, then cell culture efficiency is improved, but separation of cells from cell support residues becomes difficult
Solution Approach 1:
The patent applies fluorescent labeling to cell support residues, causing them to emit characteristic fluorescence signals that differ from cells. This enables easy differentiation and separation of cells from residues using fluorescence-activated cell sorting (FACS) or fluorescence microscopy, resolving the separation difficulty while maintaining culture efficiency
Solution Approach 2:
The patent introduces fluorescently labeled lectins as intermediaries that specifically bind to carbohydrate moieties on cell support residues. These lectins act as mediators that attach fluorescent markers to residues without affecting live cells, enabling subsequent easy separation based on fluorescence signals
2Measurement precision
If cell support residues are not separated, then measurement accuracy of cell support-derived components is improved, but contamination of cell samples occurs
Solution Approach 1:
The patent uses fluorescent labeling to give cell support residues a distinct optical signature. This allows researchers to visually identify and measure residues separately from cells using fluorescence detection, achieving accurate measurement of cell support-derived components while monitoring for contamination
Solution Approach 2:
The patent implements fluorescence-based detection as a feedback mechanism to monitor the presence and quantity of cell support residues in cell samples. This real-time feedback enables researchers to assess contamination levels and adjust separation protocols accordingly, ensuring measurement 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 simple and sensitive measurement of cell support-derived components, facilitating accurate separation and evaluation of cell cultures, particularly when using dissolvable polysaccharide cell supports.
Implementation Method 1
an aromatic compound that can produce a fluorescent or light-absorbing compound as a result of a reaction with a saccharide under heating, wherein the aromatic compound has at least one functional group selected from the group consisting of a hydroxyl group, an amino group, a nitro group and a carbonyl group
Implementation Method 2
subjecting a reaction product in the mixture after the heating to fluorescence analysis or absorption analysis
Data Source
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AI summary
Provided is a method for evaluating a sample, the method including using a mixture containing a sample including at least one member selected from the group consisting of cell support-derived components, a cell suspension containing cells, an evaluation sample obtained from a cell suspension, a sample containing a liquid and microcarriers for use in cell culture, and a sample containing a liquid obtained following treatment of microcarriers, together with at least one substance selected from the group consisting of an aromatic compound having at least one functional group selected from the group consisting of a hydroxyl group, an amino group, a nitro group and a carbonyl group, and a fluorescent dye.