Light-Chain-Only Binding Proteins for Antibody Diversity
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing antibodies lack sufficient diversity in binding proteins, necessitating the development of compositions and methods to enhance diversity and provide alternative sources of immunoglobulin-based binding proteins.
Innovation Solution
Genetically modified mice are developed with an immunoglobulin heavy chain locus replaced by light chain variable region 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 chain domains, to create enhanced diversity and specificity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If traditional antibodies with both VH and VL domains are used, then binding specificity is achieved, but diversity of binding proteins is insufficient
Solution Approach 1:
The patent extracts the VH domain from the traditional antibody structure, creating binding proteins that consist only of VL domains fused to constant regions. This extraction of the variable heavy chain domain allows for enhanced diversity by using only light chain variable regions, which have their own diverse repertoire, while simplifying the overall protein structure.
Solution Approach 2:
Instead of using the conventional VH-VL pairing approach, the patent inverts the strategy by creating binding proteins that rely solely on VL domains for antigen recognition. This inversion allows exploration of light chain-only binding specificities and expands the diversity of binding proteins beyond traditional heavy chain limitations.
2Adaptability or versatility
If VL domains are used without cognate VH domains, then new sources of diversity are created, but binding affinity may be reduced
Solution Approach 1:
The patent changes the structural parameters of the binding protein by fusing VL domains to constant regions (CH or CL) in different configurations. This parameter change allows VL domains to function independently for antigen binding while maintaining stability through constant region fusion, thereby preserving binding affinity while achieving diversity.
Solution Approach 2:
The patent creates composite binding proteins by combining VL domains with constant regions in novel fusion configurations. These composite structures integrate the diversity-capable VL domains with stability-providing constant regions, achieving both high diversity and maintained binding affinity in a single protein entity.
3Adaptability or versatility
If genetically modified mice with replaced heavy chain locus are used, then enhanced diversity of binding proteins is achieved, but manufacturing complexity increases
Solution Approach 1:
The patent segments the immunoglobulin locus into separate functional components by replacing the heavy chain variable region locus with light chain variable region loci. This segmentation allows independent manipulation and diversification of light chain sequences while maintaining separate constant region functions, thereby achieving diversity through modular genetic architecture.
Solution Approach 2:
The patent adds a new dimension to antibody diversity by creating a third locus type (light chain variable region locus in the heavy chain position) that can recombine with both heavy and light chain constant regions. This dimensional addition to the traditional VH-CL and VL-CH pairing system expands the combinatorial diversity space available for generating binding proteins.
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.