Structure-Based Antibody Humanization Method
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Solution Overview
Problem
Current methods for humanizing non-human antibodies often result in reduced antigen binding affinity and immunogenicity, as they fail to accurately predict and address the structural changes required to maintain the native conformation and function of the antibody binding site.
Innovation Solution
A structure-based approach that quantifies the topological and structural differences between non-human and human antibody sequences, allowing for specific amino acid mutations to be identified and introduced to minimize disruption to the antigen binding site, thereby enhancing the binding affinity and humanization of antibodies.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If conventional CDR grafting methods are used to humanize non-human antibodies, then the humanization level is improved, but the antigen binding affinity deteriorates
Solution Approach 1:
The method segments the antibody variable domain into CDRs and framework regions, and further segments the framework regions into sub-regions with different functional roles. This allows selective modification of framework residues that do not compromise CDR conformation or antigen binding, thereby improving humanization while maintaining binding affinity.
Solution Approach 2:
The method applies different modification strategies to different local regions of the antibody structure. Framework residues are evaluated individually based on their local structural context, solvent accessibility, and interaction with CDRs. Only residues that can be modified without affecting local conformation are changed, preserving antigen binding while improving humanization.
2Adaptability or versatility
If more amino acid residues are mutated to achieve higher humanization, then the humanization level is improved, but the structural stability deteriorates
Solution Approach 1:
The method performs preliminary computational analysis to predict the structural impact of each potential mutation before actually introducing it. Framework residues are evaluated for their role in maintaining CDR conformation and overall structural stability. Only mutations predicted to be structurally acceptable are introduced, preventing destabilization while achieving high humanization.
Solution Approach 2:
The method uses iterative refinement where each mutation is evaluated based on its impact on structural stability and CDR conformation. Computational modeling provides feedback on whether a mutation is acceptable, and this information guides subsequent mutation decisions. This feedback loop ensures that humanization proceeds without compromising structural integrity.
Data Source
AI summary
Herein is reported a method for the humanization of non-human antibodies using a structure-based scoring matrix. With the scoring matrix it is possible to determine (the requirement for and) the suitability of specific (back)mutations of amino acid residues at defined positions of a selected human germline sequence. The scoring matrix takes into account the topology, the three-dimensional structure and the interactions of the respective residue and change. Thereby the influence on antigen binding of a specific amino acid residue change can be determined.