VL Binding Proteins From Engineered Mice for Antibody Diversity
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Solution Overview
Problem
Existing antibodies lack sufficient diversity in binding proteins, necessitating the development of compositions and methods to enhance diversity in immunoglobulin-based binding proteins, particularly through the use of immunoglobulin variable regions and light chain variable domains.
Innovation Solution
Genetically modified mice are created with an immunoglobulin heavy chain locus that replaces VH, DH, and JH gene segments with human light chain variable (VL) and joining (JL) gene segments, allowing for the production of binding proteins comprising immunoglobulin variable domains from kappa (κ) and/or lambda (λ) light chains without full-length heavy chains, enhancing diversity and affinity for antigen binding.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If traditional antibody structures with VH and VL domains are used, then the binding proteins can be produced, but the diversity of binding proteins is insufficient
Solution Approach 1:
The patent segments the traditional antibody structure by separating the variable domain functions into distinct light chain variable domains (VL) that can independently bind antigens. By using only VL domains from kappa and lambda light chains without requiring cognate VH domains, the system creates modular binding units that can be combined with various constant regions to generate diverse binding proteins with different functionalities and affinities.
Solution Approach 2:
The patent applies universality by creating a platform where a single type of variable domain (VL from light chains) can perform multiple binding functions when combined with different constant regions. The VL domains can be paired with various heavy chain constant regions (CH1, CH2, CH3) and light chain constant regions (CL) to produce multiple antibody formats (IgG, IgA, IgM, etc.) from the same variable domain repertoire, greatly expanding functional diversity.
2Adaptability or versatility
If VH domains are used to enhance binding diversity, then more binding specificities can be achieved, but the affinity and specificity control becomes more difficult
Solution Approach 1:
The patent applies local quality by focusing the diversity-generating capability specifically on the light chain variable domains while maintaining controlled constant regions. The VL domains serve as the specialized binding elements with tailored affinities, while the constant regions provide standardized structural and functional properties. This localization of variability allows precise control over binding characteristics without the complexity of coordinating VH-VL pairings.
3Stability of the object's composition
If full-length heavy chains are used in binding proteins, then the structural stability is maintained, but the diversity of light chain variable domains cannot be fully exploited
Solution Approach 1:
The patent extracts the essential antigen-binding function from the traditional antibody structure by isolating the light chain variable domains and removing the requirement for heavy chain variable domains. By taking out the VH domain requirement and using only VL domains paired with constant regions, the system exploits the full diversity of light chain variable domains while maintaining structural stability through the constant region framework.
Data Source
AI summary
Genetically modified mice and methods for making an using them are provided, wherein the mice comprise a replacement of all or substantially all immunoglobulin heavy chain V gene segments, D gene segments, and J gene segments with at least one light chain V gene segment and at least one light chain J gene segment. Mice that make binding proteins that comprise a light chain variable domain operably linked to a heavy chain constant region are provided. Binding proteins that contain an immunoglobulin light chain variable domain, including a somatically hypermutated light chain variable domain, fused with a heavy chain constant region, are provided. Modified cells, embryos, and mice that encode sequences for making the binding proteins are provided.