CH3 Domain Interface Engineering for Antibody Stability

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Solution Overview

Problem

The commercial production of antibodies and Fc-containing molecules is complicated by heterogeneity, leading to decreased stability due to degradation and aggregation, which results in reduced yield and increased production costs.

Innovation Solution

Altering the CH3 domain interface of IgG1 Fc-containing proteins by substituting Ser364 with alanine or valine reduces aggregation, thereby increasing the stability and homogeneity of the final purified material.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If wild-type CH3 domain is used in Fc-containing proteins, then natural structure and function are maintained, but aggregation occurs and stability decreases

Engineering Contradiction:

Improvestability of purified polypeptideVSAvoidaggregation

Core Design Contradiction:

ReliabilityVSObject-generated harmful factors

Solution Approach 1:

The patent applies parameter changes by substituting specific amino acid residues in the CH3 domain interface (particularly Ser364, Thr366, and Ser370 positions) to alter the physical-chemical properties of the protein interface. These substitutions change the aggregation-prone parameters while maintaining the essential Fc domain structure and function, thereby reducing aggregation and improving stability of the purified polypeptide.

Inventive Principle:

Principle #35Parameter changes

2Productivity

If Fc-containing molecules are produced commercially, then therapeutic protein supply is increased, but heterogeneity leads to decreased yield and increased costs

Engineering Contradiction:

Improveyield of Fc-containing moleculesVSAvoidhomogeneity of final product

Core Design Contradiction:

ProductivityVSManufacturing precision

Solution Approach 1:

The patent modifies the CH3 domain interface parameters through amino acid substitutions to create Fc-containing molecules with improved homogeneity. This reduces degradation and aggregation during commercial production, thereby increasing yield and reducing manufacturing costs while maintaining production scale.

Inventive Principle:

Principle #35Parameter changes

3Reliability

If production processes and formulations are optimized to increase stability, then degradation and aggregation are reduced, but production complexity and costs increase

Engineering Contradiction:

Improvestability of Fc-containing moleculesVSAvoidcomplexity of production process

Core Design Contradiction:

ReliabilityVSDevice complexity

Solution Approach 1:

The patent implements preliminary action by pre-modifying the CH3 domain interface through amino acid substitutions before production. This proactive structural modification prevents degradation and aggregation during production and storage, eliminating the need for complex post-production optimization processes and reducing overall production complexity.

Inventive Principle:

Principle #10Preliminary action

Data Source

PatentEP2451840B1Design of stable and aggregation free antibody FC molecules through ch3 domain interface engineering

Publication Date: 2018.12.26 AMGEN INC

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AI summary

The present invention relates to methods of increasing stability and reducing aggregation in compositions comprising antibody Fc molecules and to composition comprising such molecules. Certain amino acid substitutions in the CH3 domain result in increased stability and reduced aggregation of compositions containing polypeptides comprising a CH3 domain, e.g., an antibody or Fc-fusion protein.