FcRn-Binding Polypeptides for EV Purification and Half-Life Extension
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Solution Overview
Problem
Current methods for isolating and purifying extracellular vesicles (EVs) face challenges such as low scalability and inefficiency in industrial production, particularly in maintaining their half-life in circulation, which hampers their therapeutic potential.
Innovation Solution
Development of FcRn binding polypeptides with a transmembrane domain that specifically bind to the FcRn receptor, allowing for affinity chromatography-based purification at physiological pH conditions, enhancing the half-life and therapeutic potential of EVs.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional isolation and purification methods are used for extracellular vesicles, then the process is simple to perform, but the scalability is low and purification efficiency is poor
Solution Approach 1:
The patent modifies the Fc domain parameters to create monomeric variants with altered binding characteristics to FcRn, enabling pH-dependent binding that facilitates purification. This parameter change in the protein structure allows for improved purification efficiency while maintaining methodological simplicity through affinity chromatography
Solution Approach 2:
The patent introduces FcRn as an intermediary binding partner in the purification process. By engineering EVs to express Fc domains that bind to FcRn, the system creates a specific interaction pathway that enables efficient capture and purification of EVs from complex biological fluids, significantly improving purification efficiency
2Reliability
If EVs are prepared for therapeutic use, then therapeutic potential is increased, but half-life in circulation is reduced
Solution Approach 1:
The patent introduces pH-dependent binding parameter changes in the Fc domain, allowing EVs to bind to FcRn at acidic pH (protection from degradation) and release at neutral pH (circulation). This parameter modification extends the half-life of EVs in circulation while preserving their therapeutic potential by protecting them from lysosomal degradation
Solution Approach 2:
The patent applies prior cushioning by engineering EVs with Fc domains that bind to FcRn before the EVs encounter degradation pathways. This pre-established binding interaction protects EVs from lysosomal degradation and extends their circulation half-life, ensuring they remain intact and therapeutically active longer
3Duration of action of moving object
If Fc domains are modified to bind FcRn, then half-life is extended, but the ability to form homodimers is lost
Solution Approach 1:
The patent applies parameter changes by modifying specific amino acid residues in the Fc domain (such as introducing charged residues at the dimerization interface) to prevent homodimer formation. This parameter modification creates monomeric Fc domains that can still bind FcRn with high affinity, extending half-life while maintaining the necessary binding stability through controlled electrostatic interactions
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 FcRn binding polypeptides enable efficient and scalable purification of EVs with improved half-life, facilitating their therapeutic and diagnostic applications by maintaining stability and circulation time.
Implementation Method 1
affinity chromatography-based purification at physiological pH conditions
Data Source
AI summary
The present disclosure pertains to polypeptides comprising a transmembrane domain and an FcRn binding site (e.g., a modified Fc domain) and nanovesicles (e.g, extracellular vesicles (EVs) and hybridosomes) comprising such polypeptides. Said polypeptides can facilitate isolation and purification of nanovesicles comprising such polypeptides. The polypeptides and nanovesicles can be used in therapeutic and/or diagnostic applications. Also provided are nucleic acids and expression vectors encoding such polypeptides as well as cells expressing said polypeptides. Further provided are methods for producing nanovesicles comprising such polypeptides and methods for purifying these nanovesicles. Compositions comprising such polypeptides or nanovesicles as well as their uses are also described.