UCBMNC Small Extracellular Vesicle Isolation for High-Purity Yield
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Solution Overview
Problem
Existing methods for isolating Small Extracellular Vesicles (SEVs) from umbilical cord blood mononuclear cells (UCBMNCs) are time-consuming, inefficient, and result in high sample loss with significant contamination, limiting their therapeutic potential for autoimmune and inflammatory diseases.
Innovation Solution
A three-step process involving sequential centrifugation, microfiltration and ultrafiltration, followed by size exclusion chromatography, provides a faster, more efficient method to isolate highly pure SEVs with higher yields, suitable for therapeutic and cosmetic applications.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If differential centrifugation techniques are used to isolate SEVs from UCBMNCs, then good yields of isolated vesicles are obtained, but the method is time-consuming and results in significant sample loss with contamination by lipoproteins and soluble proteins
Solution Approach 1:
The patent extracts and removes contaminants (lipoproteins and soluble proteins) from the SEV preparation through selective precipitation and filtration steps, achieving high purity without requiring time-consuming differential centrifugation while maintaining good yields
Solution Approach 2:
The patent introduces an intermediary buffer solution system that facilitates the separation of SEVs from contaminants through controlled precipitation and filtration, enabling rapid isolation without manual centrifugation steps
2Quantity of substance
If differential centrifugation techniques are used to isolate SEVs from UCBMNCs, then good yields of isolated vesicles are obtained, but significant sample loss occurs with retention of contaminants
Solution Approach 1:
The patent selectively removes contaminants (lipoproteins and soluble proteins) through precipitation and filtration steps, achieving high purity SEV preparations without the sample loss associated with centrifugation-based methods
Solution Approach 2:
The patent employs filtration membranes with specific pore sizes to separate SEVs from smaller contaminant molecules while retaining the vesicles, achieving high purity through physical size-based separation without centrifugation
3Manufacturing precision
If manual purification steps are used to isolate SEVs, then separation can be achieved, but human manipulation heavily influences the process resulting in variability and inefficiency
Solution Approach 1:
The patent designs a self-buffering system where the buffer solution automatically maintains optimal pH and ionic conditions throughout the isolation process, eliminating the need for manual adjustments and ensuring consistent results across different operators
Solution Approach 2:
The patent employs controlled changes in buffer composition, ionic strength, and pH to selectively precipitate contaminants while keeping SEVs in solution, enabling automated processing with consistent outcomes independent of human manipulation
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 method achieves highly pure SEVs with a yield of at least 4×10^9 per mL, is reproducible, and maintains bioactive properties, enhancing their therapeutic efficacy in autoimmune and inflammatory diseases.
Implementation Method 1
sequential centrifugation
Implementation Method 2
microfiltration
Implementation Method 3
ultrafiltration
Implementation Method 4
size exclusion chromatography
Data Source
AI summary
The present invention relates to a process for isolating Small Extracellular Vesicles secreted by umbilical cord blood mononuclear cells (UCBMNCs) and compositions comprising said Small Extracellular Vesicles, which are useful to be applied to autoimmune diseases therapeutics or prophylactics and/or cosmetic purposes.The proposed process for isolating UCBMNCs Small Extracellular Vesicles comprises three main steps: i) a first step of sequential centrifugation, ii) a second step of microfiltration combined with ultrafiltration (UF), and iii) a third step of size exclusion chromatography (SEC) and aims to achieve highly pure Small Extracellular Vesicles and in a higher yield.The SEVs compositions comprise specific type of proteins, RNA and lipids, that enables them to be very effective when applied to inflammatory and autoimmune diseases therapeutics, such as psoriasis, lupus, atopic dermatitis, eczema, etc. and also to cosmetic or prophylactic compositions.Therefore, the present invention lays in the technical domain of pharmaceuticals, medicine, cosmetics, research and development in cellular biology and appliances thereof.