Powdered Bovine Milk Exosomes for Membrane-Stable Isolation
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Solution Overview
Problem
Existing methods for isolating bovine milk exosomes often damage the exosome membrane, leading to the degradation of bioactive agents such as miRNA, which are unstable and lose their functionality quickly.
Innovation Solution
A method involving gentle centrifugation, microfiltration, and freeze-drying is used to isolate and preserve intact bovine milk exosomes, maintaining the integrity of their bilayer membrane and contents.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If conventional isolation methods are used to obtain exosomes from bovine milk, then exosomes can be isolated, but the exosome membrane is damaged and bioactive agents degrade
Solution Approach 1:
The patent modifies isolation parameters by using gentler centrifugation speeds and temperatures, adjusting pH levels, and employing specific buffer compositions to maintain exosome membrane integrity while achieving effective isolation from bovine milk
Solution Approach 2:
The patent introduces intermediary substances such as protective buffers and stabilizing agents during the isolation process that prevent membrane damage while allowing exosomes to be separated from milk components
2Productivity
If exosomes are processed or stored for extended periods, then they can be prepared for use, but the exosome membrane is damaged and bioactivity is lost
Solution Approach 1:
The patent applies preliminary protective measures by pre-cooling all equipment and reagents to low temperatures before processing, and by adding stabilizing agents at the beginning of the isolation protocol to prevent membrane damage during subsequent processing steps
Solution Approach 2:
The patent utilizes controlled phase transitions, specifically freeze-drying (lyophilization), to preserve exosomes in a stable powdered form that maintains membrane integrity and bioactivity during storage while enabling long-term preservation without refrigeration
3Loss of substance
If bioactive agents like miRNA are exposed to damage conditions, then they can be released from exosomes, but they degrade within seconds and lose functionality
Solution Approach 1:
The patent employs beforehand cushioning by incorporating protective agents in the isolation buffer that cushion and protect bioactive agents from degradation during the isolation process, preventing rapid degradation that would otherwise occur within seconds of membrane damage
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 produces powdered exosomes with high stability, retaining bioactive agents like miRNA, which can be incorporated into nutritional compositions to provide therapeutic benefits for conditions like insulin resistance and diabetes.
Implementation Method 1
centrifuging bovine milk to form a lipid fraction top layer, a whey fraction middle layer, and a first pellet of cells and debris
Implementation Method 2
The exosome-containing pellet is freeze-dried to produce the powdered exosomes
Data Source
Figure 1A~1D
Figure 1E~1I
Figure 2A
AI summary
Bovine milk-isolated powdered exosomes comprise intact exosomes. Nutritional compositions comprise protein, carbohydrate, and/or fat, and exosomes isolated from bovine milk. A method of preparing powdered exosomes comprises centrifuging bovine milk to form a lipid fraction top layer, a whey fraction middle layer, and a first pellet of cells and debris, separating the whey fraction and centrifuging the separated whey fraction to remove additional fat, casein aggregates and debris and form a substantially clear whey fraction, microfiltering the substantially clear whey fraction to remove residual debris, centrifuging the microfiltered whey fraction to obtain a pellet containing exosomes, incubating the exosome-containing pellet in aqueous medium to dissolve the pellet without disrupting exosome membranes to provide an exosome suspension, and drying the suspension to obtain the powdered exosomes. Methods of lowering a risk of developing, or treating, insulin resistance, prediabetes, or diabetes in a subject employ exosomes isolated from bovine milk.