AAV Capsid Chromatography Ligand for Full-Empty Separation

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Solution Overview

Problem

Current chromatography materials and methods struggle to efficiently and scalably separate fully packaged adeno-associated virus (AAV) capsids from not fully packaged capsids, particularly for AAV serotypes like AAV2, AAV5, AAV8, and AAV9, due to capsid diversity and small differences in isoelectric points, leading to challenges in achieving accurate quantitation and mass balance.

Innovation Solution

A chromatography device with a specific ligand structure (Formula I) and a volume of 0.1-2 mL is used for analytic separation, enabling improved resolution between fully and not fully packaged AAV capsids, allowing for scalable purification and application across various AAV serotypes.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If conventional chromatography materials and methods are used, then the separation process can be performed, but the resolution between fully packaged and not fully packaged capsids is insufficient due to small differences in isoelectric points

Engineering Contradiction:

Improveresolution between fully packaged and not fully packaged capsidsVSAvoidcapsid diversity requiring extensive optimization

Core Design Contradiction:

Measurement precisionVSDevice complexity

Solution Approach 1:

The patent modifies the chromatography ligand structure by introducing specific chemical groups (carboxymethyl and hydroxyl groups) to change the interaction parameters between the chromatography material and capsids. This chemical modification enables better resolution of capsids with small isoelectric point differences without requiring extensive method optimization for each serotype.

Inventive Principle:

Principle #35Parameter changes

2Reliability

If ultracentrifugation is used for purification, then efficient separation can be achieved, but the method is not scalable

Engineering Contradiction:

Improveseparation efficiencyVSAvoidscalability

Core Design Contradiction:

ReliabilityVSProductivity

Solution Approach 1:

The patent replaces the mechanical ultracentrifugation system with a chromatography-based separation system. This substitution maintains high separation efficiency while enabling scalability through standard chromatography equipment and procedures that can be easily scaled from analytical to preparative and manufacturing scales.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

3Reliability

If multiple filtration and chromatography steps are used, then AAV particles can be separated from cell cultures, but the process becomes complex and time-consuming

Engineering Contradiction:

Improvepurification effectivenessVSAvoidpurification process time

Core Design Contradiction:

ReliabilityVSLoss of time

Solution Approach 1:

The patent combines multiple purification functions into a single chromatography step using the modified ligand structure. The new ligand enables simultaneous separation of AAV particles from cell cultures and differentiation between fully packaged and not fully packaged capsids in one operation, reducing the number of steps and overall processing time.

Inventive Principle:

Principle #5Merging (Combining)

4Measurement precision

If conventional chromatography materials are used, then separation can be performed, but accurate quantitation and mass balance are difficult to achieve

Engineering Contradiction:

Improvequantitation accuracyVSAvoidmass balance determination

Core Design Contradiction:

Measurement precisionVSEase of operation

Solution Approach 1:

The patent introduces specific functional groups (carboxymethyl and hydroxyl groups) at specific positions on the ligand structure to create localized interaction sites that provide differential binding characteristics. This local modification enables better separation and more accurate quantitation of different capsid types, improving mass balance determination.

Inventive Principle:

Principle #3Local quality

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 device achieves high resolution and scalability, ensuring at least 80% of the AAV capsids in the eluate are fully packaged, with a capacity to process 10^10 to 10^13 capsids, and is applicable to AAV serotypes 1, 2, 4, 5, 6, 7, 8, 9, and 10, enhancing therapeutic efficacy by enriching full capsids.

Implementation Method 1

a chromatography device for use in an analytic separation method, comprising a chromatography material, which comprises a support and a ligand

Methodology Applied

Scientific EffectChromatography: Chromatography

Data Source

PatentUS20260103685A1A chromatography device, system, and use thereof for analytic separation

Publication Date: 2026.04.16 CYTIVA BIOPROCESS R&D AB

AI summary

The present disclosure is directed to a chromatography device for use in an analytic separation method, comprising a chromatography material, which comprises a support and a ligand, wherein the ligand of the chromatography material is defined by the following Formula (I) wherein R1 is selected from C1-C3 alkyl, and R2 and R3 are independently selected from C1-C3 alkyl, CH2OH, and CH2CHOHCH3, wherein the volume of the chromatography material is from about 0.1 mL to about 2 mL. Further disclosed are uses of said chromatography device and a method for separating adeno-associated virus capsids fully packaged with genetic material from adeno-associated virus capsids not fully packaged with genetic material, as well as a kit of parts and a chromatography system for use in analytic separation of adeno-associated capsids.