Ferritin Encapsulation via Flow Micromixing to Prevent Aggregation

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Solution Overview

Problem

Existing methods for encapsulating bioactive substances like siRNA and DNA in ferritin supramolecular structures face limitations due to molecular size constraints and high aggregation risks during reassembly, leading to reduced yield and increased costs.

Innovation Solution

A method using a flow micro mixer (FMM) to reassemble ferritin subunits with bioactive substances, optimizing pH conditions and flow rates to suppress misassembly and enhance encapsulation efficiency.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Ease of manufacture

If ferritin is reassembled using a batch method after disassembly, then the process allows for mixing with bioactive substances, but ferritin aggregation occurs during reassembly, reducing yield and quality

Engineering Contradiction:
Improveencapsulation processVSAvoidferritin assembly quality
Core Design Contradiction:
Ease of manufactureVSManufacturing precision

Solution Approach 1:

The patent replaces the conventional batch mixing method with a flow micro mixer system. This substitution transitions from static batch processing to dynamic flow-based mixing, enabling precise control over reassembly conditions and preventing aggregation through continuous flow dynamics and controlled shear forces.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent utilizes pH parameter changes to control ferritin disassembly and reassembly. By adjusting pH to acidic or basic conditions for disassembly, then returning to neutral pH for reassembly in the presence of bioactive substances, the process achieves controlled transformation while preventing unwanted aggregation through buffered flow conditions.

Inventive Principle:
Principle #35Parameter changes

2Productivity

If conventional batch reassembly is used, then ferritin can be reformed after disassembly, but the yield decreases due to aggregation, increasing production costs

Engineering Contradiction:
Improveencapsulation yieldVSAvoidferritin aggregation loss
Core Design Contradiction:
ProductivityVSLoss of substance

Solution Approach 1:

The flow micro mixer replaces batch mechanical stirring with controlled flow dynamics. The continuous flow system maintains ferritin subunits in suspension with controlled collision frequency and energy, preventing aggregation while promoting proper assembly, thereby increasing yield and reducing substance loss.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent implements continuous flow processing instead of intermittent batch processing. The continuous flow ensures constant renewal of reassembly conditions, preventing local accumulation of ferritin that could lead to aggregation, and maintains optimal encapsulation efficiency throughout the process.

Inventive Principle:
Principle #20Continuity of useful action

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 achieves high-yield encapsulation of bioactive substances in ferritin supramolecular structures while minimizing denaturation and aggregation, improving production efficiency.

Implementation Method 1

bringing a subunit of a protein, which forms a supramolecular structure, a bioactive substance, and a solution for forming the protein having the supramolecular structure from the subunit into contact with one another in a flow micro mixer

Methodology Applied
Scientific EffectFluid flow mixing: Convection

Implementation Method 2

the subunit is obtained by (II-1) changing pH of a solution containing the protein having the supramolecular structure to an acidic property or a basic property

Methodology Applied
Scientific EffectpH-induced disassembly: Chemical Bonding

Data Source

PatentEP4303227B1Method for producing protein having supramolecular structure in which bioactive substance is encapsulated
Publication Date: 2025.12.31 AJINOMOTO CO INC
  • EP4303227B1 patent drawingFigure 1~2
  • EP4303227B1 patent drawingFigure 3

AI summary

the present invention provides a method for producing a protein having a supramolecular structure in which a bioactive substance is encapsulated, comprising: (I) bringing a subunit of a protein, which forms a supramolecular structure, a bioactive substance, and a solution for forming the protein having the supramolecular structure from the subunit into contact with one another in a flow micro mixer.