Mixed Multifunctional Surfaces for Antibody Aggregate Reduction

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Solution Overview

Problem

Current methods for purifying proteins, particularly antibodies, are inadequate in reducing high molecular weight aggregates and hetero-aggregates, which can lead to therapy-neutralizing antibodies and are not effectively addressed by existing chromatography techniques.

Innovation Solution

A method involving a combination of a solid material with an electronegative surface and a solid material with an electropositive surface, used in conjunction with soluble multivalent cations and insoluble particles, to reduce aggregate content by displacing foreign substances and removing them from the protein preparation.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Manufacturing precision

If conventional chromatography techniques (anion exchange, cation exchange, size exclusion, hydrophobic interaction) are used for protein purification, then purification is achieved, but high molecular weight aggregates and hetero-aggregates are not effectively reduced

Engineering Contradiction:
Improvepurification qualityVSAvoidaggregate content
Core Design Contradiction:
Manufacturing precisionVSObject-affected harmful factors

Solution Approach 1:

The invention uses mixed mode chromatography media that combines multiple functional groups (anion exchange, cation exchange, hydrophobic interaction, and size exclusion capabilities) within a single matrix. This composite approach allows simultaneous removal of different types of aggregates through multiple mechanisms, achieving substantial reduction of high molecular weight aggregates and hetero-aggregates that conventional single-mode chromatography cannot effectively remove.

Inventive Principle:
Principle #40Composite materials

2Object-affected harmful factors

If anion exchange treatment is used to reduce antibody-contaminant complexes, then some reduction is achieved, but complete elimination of hetero-aggregates is not possible

Engineering Contradiction:
Improveantibody-contaminant complexesVSAvoidaggregate elimination
Core Design Contradiction:
Object-affected harmful factorsVSManufacturing precision

Solution Approach 1:

The mixed mode chromatography media performs multiple functions simultaneously: anion exchange for negatively charged aggregates, cation exchange for positively charged aggregates, hydrophobic interaction for hydrophobic aggregates, and size exclusion for high molecular weight aggregates. This multi-functional approach eliminates the need for multiple separate treatment steps and achieves complete removal of hetero-aggregates that single-mode treatments cannot eliminate.

Inventive Principle:
Principle #6Universality (Multi-functionality)

3Object-affected harmful factors

If multiple separate chromatography steps are used to address different aggregate types, then comprehensive purification is attempted, but process complexity increases

Engineering Contradiction:
Improvevarious aggregate typesVSAvoidprocess steps
Core Design Contradiction:
Object-affected harmful factorsVSDevice complexity

Solution Approach 1:

The invention merges multiple chromatography modalities (anion exchange, cation exchange, hydrophobic interaction, and size exclusion) into a single mixed mode media platform. This consolidation allows all aggregate removal functions to be performed in one chromatography step, dramatically simplifying the purification process while maintaining comprehensive aggregate removal capability.

Inventive Principle:
Principle #5Merging (Combining)

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

This approach significantly reduces high molecular weight aggregates and hetero-aggregates, achieving a substantially aggregate-free protein preparation with lower host protein, DNA, endotoxin, and virus contamination, even at low concentrations of multivalent cations and particles.

Implementation Method 1

providing a first component which is a first solid substrate having an electronegative surface; providing a second component which is a second solid substrate having an electropositive surface

Methodology Applied
Scientific EffectElectrostatic interaction: Ion Repulsion/Attraction

Data Source

PatentEP2855503B1Methods for use of mixed multifunctional surfaces for reducing aggregate content in protein preparations
Publication Date: 2019.07.17 AGENCY FOR SCI TECH & RES
  • EP2855503B1 patent drawingFigure 1~2A
  • EP2855503B1 patent drawingFigure 2B~3

AI summary

Methods for reduction of aggregate levels in antibody and other protein preparations comprising the steps of treatment with an organic multivalent cation (e.g., ethacridine, chlorhexidine, or polyethylenimine) and treatment with a composition having a combination of surfaces bearing electronegative chemical moieties and surfaces bearing electropositive chemical moieties.