Extracellular Vesicle Loading With Single-Pass Membrane Scaffolds

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Solution Overview

Problem

Current methods for loading therapeutic cargo molecules into extracellular vesicles (EVs) face challenges in achieving high efficiency and flexibility for intracellular delivery, particularly due to the limitations of multipass transmembrane proteins like CD63, which hinder the addition of targeting moieties and endosomal escape.

Innovation Solution

The use of a single-pass transmembrane protein, such as PTTG1IP, fused to the surface of EVs, allows for improved cargo loading and delivery, enabling co-expression of additional constructs and endosomal escape moieties, with enhanced efficiency compared to existing proteins like Lamp2B.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Quantity of substance

If multipass transmembrane proteins like CD63 are used for cargo loading, then loading capacity is high, but flexibility for adding targeting moieties and endosomal escape is limited

Engineering Contradiction:

Improvecargo loading capacityVSAvoidflexibility for adding targeting moieties

Core Design Contradiction:

Quantity of substanceVSAdaptability or versatility

Solution Approach 1:

The patent segments the transmembrane protein function into separate components: a scaffold protein (PTTG1IP) that provides structural support and cargo loading capacity, and separate functional modules (targeting moieties, endosomal escape peptides) that can be independently added to the extracellular domain. This segmentation allows each component to be optimized independently while maintaining overall functionality.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The patent uses an intermediary approach by employing a modular fusion protein system where the scaffold protein serves as a platform that can accommodate various functional domains. The single-pass transmembrane protein architecture acts as an intermediary structure that bridges the intracellular cargo loading machinery with extracellular targeting and escape functions.

Inventive Principle:

Principle #24Intermediary (Mediator)

2Productivity

If multipass transmembrane proteins are used, then cargo loading is efficient, but engineering of extracellular domains is hindered

Engineering Contradiction:

Improvecargo loading efficiencyVSAvoidengineering of extracellular domains

Core Design Contradiction:

ProductivityVSEase of manufacture

Solution Approach 1:

The patent inverts the traditional multipass transmembrane protein architecture to a single-pass configuration. This inversion allows the extracellular domain to be accessible and engineerable from the outside, while the intracellular domain maintains the cargo loading function. The single-pass architecture reverses the accessibility constraint, making both domains engineerable without the topological restrictions of multipass proteins.

Inventive Principle:

Principle #13The other way round (Inversion)

3Ease of manufacture

If single-pass transmembrane proteins are used, then ease of engineering is improved, but loading capacity may be reduced

Engineering Contradiction:

Improveease of engineeringVSAvoidcargo loading capacity

Core Design Contradiction:

Ease of manufactureVSQuantity of substance

Solution Approach 1:

The patent changes key parameters of the transmembrane protein architecture: switching from multipass to single-pass configuration, optimizing the transmembrane helix length and composition, and adjusting the extracellular and intracellular domain sizes. These parameter changes enable both ease of engineering and sufficient cargo loading capacity by finding the optimal balance point in the protein structure.

Inventive Principle:

Principle #35Parameter changes

Data Source

PatentUS20260034239A1Loaded extracellular vesicle

Publication Date: 2026.02.05 OXFORD UNIVERSITY INNOVATION LTD

AI summary

The present invention relates to compositions comprising an extracellular vesicle (EV). In particular, the extracellular vesicle (EV) comprises a single pass EV transmembrane protein fused to a moiety on the surface of the EV and/or a cargo molecule. The composition may be used to deliver the cargo molecule. Methods of manufacturing the composition are also provided.