Enzyme Composition for Cell Separation
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Solution Overview
Problem
Current methods for separating cells from biological tissues face challenges in degrading structural proteins without damaging target cells, as the collagen content in tissues varies by species, age, and environment, leading to inconsistent yields and quality in cell isolation.
Innovation Solution
A method using a degrading-enzyme composition with collagenase H and collagenase G, along with a neutral protease like thermolysin, is developed, where the weight ratio of collagenase H to collagenase G is optimized based on the collagen I and III composition of the tissue, to efficiently separate cells while minimizing damage.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If a protein-degrading enzyme such as collagenase is used to degrade intercellular matrix to separate cells from biological tissue, then cell separation efficiency is improved, but the target cells may be damaged or degraded along with the intercellular matrix
Solution Approach 1:
The patent segments the protein-degrading enzyme system into multiple specific enzymes (collagenase for collagen degradation, elastase for elastin degradation, hyaluronidase for hyaluronic acid degradation). Each enzyme targets a specific component of the intercellular matrix, allowing selective degradation of structural proteins while preserving target cells through controlled, component-specific action.
Solution Approach 2:
The patent applies different enzymes with specific substrate specificities to different regions or components of the intercellular matrix. Collagenase acts on collagen fibers, elastase on elastin, and hyaluronidase on hyaluronic acid, creating localized degradation effects that collectively dissolve the matrix while minimizing non-specific damage to cells.
2Ease of operation
If a fixed composition of protein-degrading enzyme is used for cell separation, then the process is simple to operate, but the separation yield and quality vary when applied to tissues from different species, ages, or environments
Solution Approach 1:
The patent creates a universal cell separation system that functions across different tissue types, species, and conditions by combining multiple enzymes with broad substrate coverage. The enzyme composition includes collagenase, elastase, and hyaluronidase, which collectively address the diverse protein components found in various intercellular matrices, making the method universally applicable while maintaining consistent results.
Solution Approach 2:
The patent adjusts enzyme composition parameters (types and ratios of collagenase, elastase, hyaluronidase) based on the specific characteristics of the tissue being processed. By varying enzyme concentrations and combinations according to tissue source, age, and collagen content, the method adapts to different conditions while maintaining operational simplicity through a standardized multi-enzyme approach.
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 allows for stable and efficient separation of cells with high biological activity by determining the optimal enzyme composition based on tissue-specific collagen content, resulting in improved yield and quality of isolated cells.
Implementation Method 1
it is necessary to treat intercellular matrix with a protein-degrading enzyme such as collagenase
Implementation Method 2
degrading the pancreas tissue without damaging the pancreatic islets
Implementation Method 3
using a degrading-enzyme composition with collagenase H and collagenase G, along with a neutral protease like thermolysin
Data Source
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AI summary
Provided is a method for efficiently and stably separating cells having a high biological activity from a biological tissue, by using a degrading-enzyme composition, which is prepared by adding an enzyme for degrading a major protein of the biological tissue in an amount determined depending on the composition of the major protein to a predetermined amount of a neutral protease and/or a protease derived from Clostridium sp. According to this method, the type and amount of protein-degrading enzyme to be used for isolating cells can be determined from the composition of a major protein of the biological tissue. Thus, cells having a high biological activity can be efficiently separated while reducing the amount of protein-degrading enzyme to be used.