Lotus Leaf-Derived Exosome Purification via Ultrafiltration
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Solution Overview
Problem
Current methods for extracting plant-derived exosome-like nanoparticles (PDENs) from living organisms are inefficient and lack standardization, which affects the stability and bioactivity of the extracted particles.
Innovation Solution
The extraction of lotus leaf-derived exosomes using methods such as polymer precipitation, ultracentrifugation, ultrafiltration, density-gradient centrifugation, and size-exclusion chromatography, with ultrafiltration being a preferred method, to establish a standard purification method and qualitative standards for these exosomes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional exosome purification methods are used to extract plant-derived exosome-like nanoparticles, then the extraction process can be performed, but the particle quality and stability are compromised due to lack of standardization
Solution Approach 1:
The patent establishes specific purification parameters including centrifugation speed (100,000-150,000 ×g), filtration membrane pore size (0.01-0.1 μm), and pH value (7.0-9.0) to standardize the extraction process. These controlled parameter changes ensure consistent particle quality and stability across different extractions, resolving the contradiction between reliability and ease of manufacture.
Solution Approach 2:
The purification process is divided into multiple sequential steps: low-speed centrifugation to remove debris, ultrafiltration through specific membrane pores, high-speed centrifugation to separate exosomes from other components, and pH adjustment. This segmentation allows each step to optimize for specific purification goals, achieving standardized high-quality extraction.
2Quantity of substance
If multiple extraction methods are applied to obtain sufficient exosomes, then the particle yield increases, but the process complexity and time consumption increase
Solution Approach 1:
The patent employs continuous centrifugation at optimized speeds (100,000-150,000 ×g) and continuous filtration through ultrafiltration membranes throughout the extraction process. This continuous action maintains high particle yield while avoiding the need for multiple discrete complex steps, thus reducing overall process complexity.
Solution Approach 2:
The ultrafiltration step serves multiple functions simultaneously: it filters particles based on size (0.01-0.1 μm), concentrates the exosomes, and pre-separates them from larger debris. This multi-functionality reduces the need for separate purification steps, simplifying the overall process while maintaining high yield.
3Manufacturing precision
If aggressive purification steps are used to remove contaminants, then the particle purity increases, but the particle stability and bioactivity may be compromised
Solution Approach 1:
The patent carefully controls pH value (7.0-9.0) and temperature (4°C) parameters throughout the purification process to maintain particle stability. By adjusting these parameters gently rather than using aggressive conditions, the method achieves high purity while preserving particle integrity and bioactivity.
Solution Approach 2:
The ultrafiltration membrane acts as an intermediary that selectively separates exosomes from contaminants based on size without direct contact or harsh treatment. This gentle separation mechanism achieves high purity while maintaining particle stability, avoiding the damaging effects of aggressive purification methods.
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 established methods result in lotus leaf-derived exosomes with stable particle sizes and zeta potentials, demonstrating their potential for alleviating inflammatory responses and wound healing, and can be used in preparing anti-inflammatory or wound-healing drugs, compositions, or nutritional supplements.
Implementation Method 1
ultrafiltration, density-gradient centrifugation, and size-exclusion chromatography
Implementation Method 2
density-gradient centrifugation
Implementation Method 3
density-gradient centrifugation
Implementation Method 4
size-exclusion chromatography
Implementation Method 5
polymer precipitation
Data Source
AI summary
The present invention provides a lotus-derived extracellular vesicle (LDEV), which are extracted from lotus leaves by a group selected from the following extraction methods: polymer precipitation method, ultracentrifugation method, ultrafiltration method, density gradient centrifugation method and size-exclusion chromatography. The present invention also provides an anti-inflammatory composition containing the LDEV and the use of the LDEV. The LDEV extracted through different separation methods in the present invention have similar particle sizes and stable zeta potentials. In addition, the present invention has experimentally confirmed that the LDEV can be used to alleviate inflammatory reactions or wound healing, and can further be prepare as anti-inflammatory or wound healing drugs, compositions or nutritional supplements.