Anti-LAG-3 Antibody Deamidation Reduction via CDR2 Engineering
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Solution Overview
Problem
Therapeutic antibodies are prone to deamidation, leading to structural and biological alterations, immunogenicity, and reduced activity, which poses a significant challenge in maintaining their potency and stability during manufacturing and storage.
Innovation Solution
A modified form of the monoclonal antibody 25F7 with altered critical binding regions, specifically the heavy chain CDR2 domain, exhibits enhanced thermal and chemical stability, reduced deamidation, and maintained high binding affinity to human LAG-3.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If therapeutic antibodies are used, then high binding affinity and functional activity are achieved, but deamidation occurs leading to reduced stability and immunogenicity
Solution Approach 1:
The patent applies parameter changes by modifying specific amino acid residues in the antibody sequence (particularly in the CDR regions and framework regions) to reduce susceptibility to deamidation. This involves changing chemical parameters of the antibody structure while maintaining its binding function, directly addressing the contradiction between stability and deamidation resistance
Solution Approach 2:
The patent converts the harmful effect of deamidation into a benefit by strategically introducing amino acid substitutions that prevent deamidation at critical sites. The modification approach transforms the vulnerability of asparagine and glutamine residues into an opportunity to enhance stability through controlled sequence engineering
2Stability of the object's composition
If amino acid modifications are made to reduce deamidation, then stability is improved, but binding affinity may be reduced
Solution Approach 1:
The patent applies local quality by making targeted amino acid modifications specifically at regions prone to deamidation (such as asparagine and glutamine residues in CDR and framework regions) while leaving the critical antigen-binding interface largely intact. This localized approach ensures stability improvement without compromising binding affinity
Solution Approach 2:
The patent carefully selects amino acid substitutions that change the chemical properties of susceptible residues (e.g., replacing asparagine with glutamine or other residues) to reduce deamidation while maintaining the overall structural and functional parameters of the antibody
3Stability of the object's composition
If storage conditions are optimized, then deamidation is reduced, but manufacturing complexity increases
Solution Approach 1:
The patent applies preliminary action by incorporating stability-enhancing amino acid modifications into the antibody sequence before production. This proactive approach prevents deamidation during manufacturing and storage, eliminating the need for complex post-manufacturing stabilization measures or specialized storage conditions
Data Source
AI summary
The present invention provides isolated monoclonal antibodies that specifically bind LAG-3, and have optimized functional properties compared to previously described anti-LAG-3 antibodies, such as antibody 25F7 (US 2011/0150892 A1). These properties include reduced deamidation sites, while still retaining high affinity binding to human LAG-3, and physical (i.e., thermal and chemical) stability. Nucleic acid molecules encoding the antibodies of the invention, expression vectors, host cells and methods for expressing the antibodies of the invention are also provided, as well as immunoconjugates, bispecific molecules and pharmaceutical compositions comprising the antibodies. The present invention also provides methods for detecting LAG-3, as well as methods for treating stimulating immune responses using an anti-LAG-3 antibody of the invention. Combination therapy, in which the antibodies are co-administered with at least one additional immunostimulatory antibody, is also provided.