Engineered Ig Lambda Light Chains for Broader Human Antibody Repertoires
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Solution Overview
Problem
Current methods for producing human monoclonal antibodies in genetically engineered animals do not maximize human antibody repertoires, necessitating improved in vivo systems for generating human monoclonal antibodies.
Innovation Solution
Engineering the germline genome of rodents to include human Vλ and Jλ gene segments operably linked to Cλ genes, while removing rodent Cκ genes, to enhance human immunoglobulin λ light chain production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If rodent Cκ genes are removed and human Vλ/Jλ gene segments are introduced at the immunoglobulin κ locus, then human antibody repertoire diversity is improved, but device complexity increases
Solution Approach 1:
The patent combines human Vλ and Jλ gene segments with rodent Cκ gene loci to create a hybrid system. This merging allows the rodent to produce human lambda light chains using the rodent constant region framework, thereby expanding human antibody repertoire while maintaining compatibility with the rodent immune system architecture.
Solution Approach 2:
The patent introduces human immunoglobulin gene segments into a non-human mammalian genome, adding a dimensional layer of cross-species genetic integration. This enables the generation of human-like antibodies in rodent systems, effectively creating a trans-dimensional genetic expression system that overcomes species-specific antibody production limitations.
2Adaptability or versatility
If human Vλ and Jλ gene segments are introduced at the immunoglobulin κ locus, then junctional diversity in light chains is improved, but manufacturing precision requirements increase
Solution Approach 1:
The patent employs pre-designed targeting vectors containing human Vλ and Jλ gene segments with defined regulatory elements and homology regions. These vectors are constructed beforehand with precise sequences to ensure correct integration at the rodent Cκ locus, thereby achieving high junctional diversity while maintaining manufacturing precision through advance preparation of genetic constructs.
Solution Approach 2:
The patent uses targeting vectors as intermediary carriers to deliver human Vλ and Jλ gene segments to the rodent Cκ locus. These vectors serve as mediators that facilitate precise integration by providing homology arms and regulatory sequences, thereby reducing the direct complexity of genome editing while ensuring accurate gene segment placement and expression.
Data Source
AI summary
Non-human animals (and/or non-human cells) and methods of using the same are provided, which non-human animals (and/or non-human cells) have a genome comprising human antibody-encoding sequences (i.e., immunoglobulin genes). Non-human animals described herein express antibodies that contain immunoglobulin (Ig) light chains characterized by the presence of human Vλ domains. Non-human animals provided herein are, in some embodiments, characterized by expression of antibodies that contain human Vλ light chains that are encoded by human Igλ light chain-encoding sequences inserted into an endogenous Igκ light chain locus of said non-human animals. Methods for producing antibodies from non-human animals are also provided, which antibodies contain human variable regions and mouse constant regions.