Stabilized Chimeric Fabs with Lambda Light Chain Modifications
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Solution Overview
Problem
Chimeric Fabs with lambda light chains often exhibit decreased thermal stability, making them difficult to manufacture and store, and existing methods fail to effectively enhance their stability without compromising antigen-binding affinity.
Innovation Solution
Introduction of stabilizing amino acid modifications in the Vlambda sequence of chimeric Fabs, specifically at residues 83, 85, 105, and 106, to increase thermal stability while maintaining antigen-binding affinity, resulting in a stabilized chimeric Fab with enhanced thermal stability compared to wild-type counterparts.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If chimeric light chains are constructed with Vlambda-Ckappa or Vkappa-Clambda configurations, then the antibody format is improved and immunogenicity is reduced, but thermal stability decreases
Solution Approach 1:
The patent applies local quality by making specific amino acid substitutions at particular positions (83, 85, 105, 106) in the Vlambda sequence while leaving the rest of the chimeric light chain structure intact. This localized modification approach improves thermal stability without compromising the overall chimeric format and antigen-binding capability.
Solution Approach 2:
The patent changes the amino acid sequence parameters at specific positions in the Vlambda domain to enhance thermal stability. By substituting specific residues (e.g., position 83, 85, 105, 106), the patent optimizes the physical-chemical properties of the chimeric light chain while maintaining its functional characteristics.
2Stability of the object's composition
If stabilizing amino acid modifications are introduced in the Vlambda sequence, then thermal stability increases, but there is a risk of affecting antigen-binding affinity
Solution Approach 1:
The patent carefully selects specific positions (83, 85, 105, 106) in the Vlambda sequence for modification, avoiding the CDR regions that are critical for antigen binding. This localized approach ensures that thermal stability is improved while antigen-binding affinity is preserved.
Solution Approach 2:
The patent introduces a limited number of stabilizing amino acid modifications at strategically chosen positions rather than attempting to optimize the entire sequence. This partial action approach is sufficient to achieve the desired thermal stability improvement without risking disruption of antigen-binding functionality.
Data Source
AI summary
Provided herein are stabilized chimeric Fabs derived from a parent chimeric Fab having a lambda light chain. The stabilized chimeric Fabs comprise an immunoglobulin heavy chain polypeptide construct from the parent chimeric Fab, having a CH1 sequence and a VH sequence, as well as a Vlambda-Ckappa chimeric light chain construct. The Vlambda sequence of the chimeric light chain construct corresponds to that of the parent chimeric Fab, and comprises one or more stabilizing amino acid modifications that increase the thermal stability of the stabilized chimeric Fab compared to the parent chimeric Fab. The stabilized Fabs are useful as therapeutic polypeptides, or can be used to prepare antibody constructs in other formats. The stabilized chimeric Fabs may also be useful generally to increase the stability of antibodies having lambda light chains.