Self-Assembling Polypeptide Nanoparticles for Oligonucleotide Delivery
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Solution Overview
Problem
Existing oligonucleotide delivery systems face challenges such as suboptimal pharmacokinetic properties, susceptibility to nucleases, urinary excretion, and immune responses, with virus-like particles being complex and costly to produce and associated with safety concerns.
Innovation Solution
Design and production of non-naturally occurring polypeptides that self-assemble into nanoparticles capable of encapsulating oligonucleotides with high binding affinity, efficiently entering mammalian cells and releasing cargo to modulate gene expression, while being safe and biodegradable.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If virus-like particles are used for oligonucleotide delivery, then delivery efficiency is improved, but production complexity and cost increase
Solution Approach 1:
The patent uses computationally designed protein sequences that copy and adapt structural motifs from viral capsids and bacterial shells to create synthetic nanocages. These designed proteins self-assemble into virus-like particles with controlled geometry (T=1, T=3, T=4 symmetries) that can deliver oligonucleotides without requiring complex viral production systems
Solution Approach 2:
The patent employs recombinant protein expression in E. coli to produce the self-assembling nanocage proteins at low cost. The proteins are expressed as inclusion bodies, purified through simple chromatography, and refolded to achieve functional assembly. This replaces expensive viral production facilities with inexpensive bacterial fermentation and purification processes
2Reliability
If oligonucleotides are delivered directly, then therapeutic effect is achieved, but susceptibility to nucleases and immune responses occurs
Solution Approach 1:
The patent encapsulates oligonucleotide therapeutics inside protein nanocage structures. The nanocages have internal cavities that bind and protect the oligonucleotides from nucleases in circulation. The cargo is nested within the protective protein shell, preventing degradation until delivery to the target cell
Solution Approach 2:
The protein nanocage acts as an intermediary carrier between the oligonucleotide therapeutic and the target cell. It protects the cargo from immune recognition and nuclease degradation in the bloodstream, while facilitating cellular uptake through receptor-mediated endocytosis. The nanocage mediates the delivery process without requiring direct administration of naked oligonucleotides
3Productivity
If self-assembling protein nanocages are used, then scalability is improved, but manufacturing precision requirements increase
Solution Approach 1:
The patent designs nanocages as assemblies of identical protein subunits that self-organize into symmetric structures (T=1 with 60 subunits, T=3 with 180 subunits, T=4 with 240 subunits). Each subunit contains standardized interface motifs that guide precise self-assembly. This modular segmentation allows scalable production through recombinant expression of individual subunit genes
Solution Approach 2:
The patent uses computational design to optimize protein sequence parameters including disulfide bridge positions, hydrophobic core packing, and interface salt bridges. These parameter optimizations ensure that the proteins fold correctly and assemble with high precision into the desired symmetric nanocage structures. The designed parameters compensate for variations in expression and purification conditions
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 nanoparticles provide a scalable, economic, and effective platform for intracellular delivery of therapeutic oligonucleotides, minimizing undesirable properties of prior art delivery vehicles and allowing controlled cargo release.
Implementation Method 1
designed and non-naturally occurring novel polypeptides that are not only capable of self-assembling into nanoparticles
Implementation Method 2
nanoparticles are furthermore capable of encapsulating negatively charged macromolecules such as oligonucleotides with high binding affinity
Data Source
AI summary
The present invention relates to polypeptides self-assembling into nanoparticles. In particular, the invention relates to a polypeptide comprising an amino acid sequence I (SEQ ID NO: 1), a nucleic acid sequence encoding said polypeptide, a nanoparticle comprising at least one polypeptide of the invention, a complex comprising said nanoparticle and one or more cargo molecules, and a method for transfecting a cell with said complex.