AAV Purification Using Surfactant and TFF

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Solution Overview

Problem

Current methods for producing and purifying adeno-associated virus (AAV) vectors are inefficient and result in low-purity, biologically inactive viral particles, posing challenges for safe and effective gene therapy.

Innovation Solution

A method involving the use of CHAPS and deoxycholic acid surfactants in combination with tangential flow filtration (TFF) to treat and purify AAV-containing samples, effectively removing hollow particles and maintaining high biological activity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Manufacturing precision

If conventional affinity purification using VHH antibody is used, then purification efficiency is improved, but biological activity is reduced due to low pH elution conditions

Engineering Contradiction:

Improvepurification efficiencyVSAvoidbiological activity

Core Design Contradiction:

Manufacturing precisionVSReliability

Solution Approach 1:

The invention changes the pH parameter from acidic (pH 2.0) to neutral (pH 7.4), eliminating the need for pH adjustment during elution while maintaining high purification efficiency through the TFF process with surfactant treatment

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The invention replaces the chemical affinity-based purification system with a physical filtration system (TFF), substituting the need for chemical elution with a mechanical filtration process that preserves biological activity

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

2Manufacturing precision

If complex conventional purification processes are used, then purification degree is improved, but process complexity and impact on biological activity increase

Engineering Contradiction:

Improvepurification degreeVSAvoidprocess complexity

Core Design Contradiction:

Manufacturing precisionVSDevice complexity

Solution Approach 1:

The invention segments the purification process into distinct functional steps: surfactant treatment to solubilize contaminants, followed by TFF for physical separation, allowing each step to be optimized independently while maintaining overall simplicity

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The invention introduces surfactant as an intermediary substance that facilitates the separation process by solubilizing hollow particles and contaminants, enabling their removal through TFF without complex chemical procedures

Inventive Principle:

Principle #24Intermediary (Mediator)

3Manufacturing precision

If conventional purification methods are used, then some purification is achieved, but hollow particle contamination remains difficult to remove

Engineering Contradiction:

Improvepurification degreeVSAvoidhollow particle contamination

Core Design Contradiction:

Manufacturing precisionVSObject-generated harmful factors

Solution Approach 1:

The invention converts the harmful hollow particles into removable contaminants by using surfactant to solubilize them, transforming them from protected viral structures into removable micellar complexes that can be easily filtered away

Inventive Principle:

Principle #22Blessing in disguise (Convert harm into benefit)

Solution Approach 2:

The invention extracts hollow particles from the viral preparation by solubilizing them with surfactant and removing them through TFF filtration, physically separating them from the intact viral particles

Inventive Principle:

Principle #2Taking out (Extraction)

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-purity AAV vectors with retained biological activity, significantly reducing hollow particle contamination, which is difficult to remove with conventional methods, thereby enhancing the safety and efficacy of gene therapy.

Implementation Method 1

have then used CHAPS, a surfactant (amphoteric surfactant) that forms amphoteric ionic micelles, in combination with deoxycholic acid, an anionic surfactant

Methodology Applied

Scientific EffectSurfactant micelle formation: Surfactant

Implementation Method 2

have then introduced a filtration operation by tangential flow filtration (TFF; alias: cross flow filtration)

Methodology Applied

Scientific EffectTangential flow filtration: Filter (physical)

Data Source

PatentUS20250197817A1Virus purification method

Publication Date: 2025.06.19 THE UNIV OF TOKYO

AI summary

It is an object of the present invention to provide a method for producing a virus, for use in obtaining viral particles with high purity, and the present method is more efficient than conventional methods. Specifically, the present invention relates to a method for obtaining a virus, comprising a step of treating a virus-containing sample with a surfactant. The surfactant may be one or more selected from the group consisting of an amphoteric surfactant, an anionic surfactant, a cationic surfactant, and a nonionic surfactant. In addition, the method for obtaining a virus according to the present invention may comprise a step of performing tangential flow filtration (TFF) to purify the virus.