Exosome Cargo Protein Loading via Photo-Specific Binding
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Solution Overview
Problem
Current methods for delivering cargo proteins into cells are inefficient, often resulting in protein misfolding, nonspecific transfer, immune reactions, high costs, and low yields, and nanoparticles used for delivery can be toxic and difficult to separate from cargo proteins, limiting their effectiveness.
Innovation Solution
The development of exosomes loaded with cargo proteins using photo-specific binding proteins, where a fusion protein composed of an exosome-specific marker and a cargo protein is expressed, allowing for efficient loading and separation of the cargo protein within the exosome, enabling targeted delivery to cytosol.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If recombinant protein with protein transduction domains is used for delivery, then the cargo protein can enter the cytosol through the cell membrane, but the protein refolding is not performed properly, the activity is decreased, the protein is nonspecifically transferred, the risk of causing an immune reaction in vivo is large, the cost is high, and the yield is low
Solution Approach 1:
The patent uses exosomes as intermediary carriers to deliver cargo proteins into cells. Instead of directly using recombinant proteins with transduction domains, the invention encapsulates the cargo protein within exosomes, which then facilitate entry into the cytosol. This intermediary approach resolves the contradiction by maintaining high delivery efficiency while avoiding the drawbacks of direct protein transduction methods.
2Reliability
If nanoparticles are used for cargo protein delivery, then the cargo protein can enter the cytosol through endocytosis, but the nanoparticles are difficult to be separated from the cargo protein and toxicity of the nanoparticles can be another problem
Solution Approach 1:
The patent changes the physical and chemical parameters of the delivery system by using exosomes - natural lipid bilayer vesicles with sizes of 30-150 nm that are biocompatible and non-toxic. Unlike synthetic nanoparticles, exosomes can be easily separated from cargo proteins after delivery and do not exhibit toxicity, thus resolving the contradiction between delivery efficiency and safety.
3Reliability
If fusion protein of PTDs and cargo protein is produced, then the cargo protein can enter the cytosol, but a separation process is required which increases cost and decreases yield
Solution Approach 1:
The patent extracts the delivery function from the cargo protein itself by using exosomes as separate delivery vehicles. Instead of creating fusion proteins that require separation processes, the invention loads cargo proteins into exosomes, which naturally facilitate cellular entry. This extraction approach eliminates the need for complex separation processes, reducing manufacturing costs and increasing yield while maintaining delivery efficiency.
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 allows for high-yield, specific delivery of cargo proteins into cells, maintaining their activity and reducing toxicity, with the potential for treating various diseases by regulating intracellular signaling and delivering therapeutic proteins effectively.
Implementation Method 1
a photo-specific binding protein, and a method for preparing exosome loaded with the cargo protein using the photo-specific binding protein
Data Source
AI summary
The present invention relates to a method for the mass-production of exosome comprising a cargo protein, a vector for preparing the exosome, exosome including a cargo protein prepared by the method, and a method for loading the cargo protein to cytosol by using the exosome prepared thereby. According to the method for preparing an exosome comprising a cargo protein provided by the present invention, the exosome loaded with a cargo protein can be produced with a high yield, so that it can be used broadly in the treatment of disease using the exosome.