Plant Exosome Isolation via Ultracentrifugation and TFF
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for isolating plant exosomes are limited in scalability, purity, and uniformity, particularly when dealing with hard flesh or peel plants, and are not suitable for large-scale commercial production due to issues like contamination and low efficiency in removing vacuoles and impurities.
Innovation Solution
A method combining ultracentrifugation and tangential-flow filtration (TFF) is used to isolate high purity plant exosomes, which allows for continuous processing, easy operation in a sterile state, and minimizes adsorption to filtration filter pores, enabling large-scale production with uniform particle size distribution.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If density gradient ultracentrifugation method using sucrose gradient is used to isolate plant exosomes, then exosome isolation can be achieved, but the method is difficult to utilize in commercial production and has low productivity
Solution Approach 1:
The patent extracts and removes the sucrose gradient step from the traditional isolation method, replacing it with a simplified ultracentrifugation protocol that achieves equivalent purification without the time-consuming density gradient layering process, thereby maintaining purity while improving productivity
Solution Approach 2:
The patent optimizes ultracentrifugation parameters (speed, time, temperature) to achieve effective exosome isolation without requiring density gradient separation, changing the physical parameters of the centrifugation process to replace the complex chemical gradient method with a more efficient physical separation approach
2Manufacturing precision
If filtration filter is used to separate exosomes from plant juices, then size-based separation can be achieved, but impurities and exosomes are adsorbed and accumulated in filter pores causing rapid lowering of filtration efficiency
Solution Approach 1:
The patent replaces the mechanical filtration system with a centrifugal separation system, substituting the physical filtering mechanism that causes pore clogging with a centrifugal force-based separation mechanism that partitions components by density and size without contact with filter pores, thereby eliminating adsorption accumulation and maintaining continuous high efficiency
Solution Approach 2:
The patent introduces an ultracentrifugation step as an intermediary separation process between raw plant juice extraction and final exosome collection, using centrifugal force as a mediator to separate exosomes from impurities without direct contact with filtration media, thus preventing the adsorption and accumulation problem
3Manufacturing precision
If conventional isolation methods are used for plant exosomes, then isolation can be achieved, but scalability and uniformity are limited particularly for hard flesh or peel plants
Solution Approach 1:
The patent segments the isolation process into distinct standardized stages (homogenization, ultracentrifugation, resuspension) that can be independently optimized and scaled, allowing the method to be adapted to different plant types including hard flesh and peel plants while maintaining uniformity through consistent procedural steps
Solution Approach 2:
The patent develops a universal isolation protocol that functions effectively across diverse plant types (soft flesh, hard flesh, peel, seeds), making the method broadly applicable and scalable while maintaining consistent exosome isolation quality through standardized ultracentrifugation parameters that work for all plant matrices
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 method effectively isolates high purity plant exosomes on a large scale, improving productivity and purity while preventing contamination, and is suitable for GMP production, facilitating process scale-up and commercialization.
Implementation Method 1
a first ultracentrifugation step to obtain a supernatant and a first pellet
Implementation Method 2
a tangential-flow filtration step to obtain a second supernatant and a second pellet
Data Source
AI summary
The present disclosure relates to a large-scale production method of plant exosomes. The method of the present disclosure can isolate high purity plant exosomes from a large amount of raw plants, using centrifugation and TFF, which can process a large amount of plant raw materials at once. This improves a conventional isolation process of plant exosomes stayed at the laboratory level, and thereby, suggests an easy process for large-scale production.