Bispecific Antibody Mutations for Lower Immunogenicity and Purity
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Solution Overview
Problem
Developing bispecific antibodies is challenging due to the difficulty in creating structures and amino acid sequences that can be produced at good yield and purity while avoiding immunogenicity, which reduces their suitability for therapeutic use.
Innovation Solution
Introduce secondary mutations to bispecific antibodies, specifically in the RUBY format, to reduce immunogenicity and improve purity by minimizing binding to MHC class II and protein A sites, thereby enhancing manufacturing efficiency and reducing unwanted immune responses.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If primary mutations are introduced to promote heavy chain-light chain association, then the association efficiency is improved, but the immunogenicity of the polypeptide increases
Solution Approach 1:
The patent applies local quality by making specific targeted mutations at defined positions (e.g., K114A in CKappa, E187K in CH1) rather than global modifications. These localized changes in the CH1 and CKappa/CLambda regions promote heavy chain-light chain association while minimizing overall immunogenicity impact by affecting only specific interaction sites.
Solution Approach 2:
The patent employs parameter changes by systematically varying amino acid substitutions at specific positions to optimize the balance between association efficiency and immunogenicity. Different mutation combinations (e.g., K114A, E187K, T114A) are tested to find the optimal parameter set that achieves stable association with acceptable immunogenicity profiles.
2Manufacturing precision
If mutations are introduced to improve manufacturing purity, then the purity during manufacture is improved, but the complexity of the polypeptide structure increases
Solution Approach 1:
The patent applies the taking out principle by removing problematic interactions through specific mutations. For example, mutations in the CKappa/CLambda and CH1 regions eliminate unwanted binding to protein A and MHC class II, extracting these harmful interactions from the polypeptide structure to improve manufacturing purity.
Solution Approach 2:
The patent uses preliminary action by incorporating purity-improving mutations into the polypeptide design before manufacturing. The mutations (e.g., in CKappa positions 114, 116 and CH1 position 187) are pre-engineered to prevent formation of unwanted complexes during production, avoiding the need for complex post-manufacturing purification steps.
3Productivity
If the polypeptide is designed for high yield production, then the manufacturing yield is improved, but the immunogenicity may increase reducing therapeutic suitability
Solution Approach 1:
The patent applies parameter changes by systematically optimizing amino acid substitutions at key positions to achieve the desired balance. The mutations (such as K114A in CKappa and E187K in CH1) are selected to maintain high expression yield while simultaneously reducing immunogenicity by altering protein A and MHC class II binding characteristics.
Solution Approach 2:
The patent uses local quality by making targeted mutations in specific regions (CKappa, CLambda, CH1) that are responsible for both yield and immunogenicity. These localized changes allow optimization of expression levels while minimizing unwanted immune responses through precise modification of interaction interfaces.
Data Source
AI summary
The present invention relates to polypeptides (in particular, bispecific antibodies) including new mutations that reduce immunogenicity and/or improve purity during manufacture.