Engineered Immunoglobulin Lambda Locus for Human Antibody Diversity
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Solution Overview
Problem
Current systems for producing human monoclonal antibodies in transgenic animals do not maximize human antibody repertoires, limiting the development of effective therapeutic antibodies.
Innovation Solution
Engineering non-human animals with an immunoglobulin lambda light chain locus that includes operably linked human Vλ, Jλ, and Cλ gene segments, along with human immunoglobulin enhancers, to enhance the diversity and expression of human Vλ regions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If transgenic animals are engineered with human antibody genetic material, then human monoclonal antibodies can be produced in vivo, but the human antibody repertoire is not maximized
Solution Approach 1:
The patent divides the immunoglobulin lambda light chain locus into distinct functional segments: human Vλ gene segments (multiple), human Jλ gene segments (multiple), human Cλ region genes (multiple), and rodent Cλ region genes. This segmentation allows each segment to contribute specifically to repertoire diversity while maintaining organized genetic structure that facilitates proper expression.
Solution Approach 2:
The engineered locus serves multiple functions simultaneously: it maintains the structural integrity required for proper immunoglobulin expression, incorporates diverse human gene segments to maximize repertoire diversity, and ensures compatible expression in the rodent host system through inclusion of both human and rodent constant region genes.
2Quantity of substance
If multiple human Vλ gene segments are incorporated into the immunoglobulin locus, then antibody repertoire diversity is enhanced, but the genetic engineering complexity increases
Solution Approach 1:
Multiple human Vλ gene segments are nested within the engineered immunoglobulin lambda light chain locus, organized in a hierarchical structure where Vλ segments are positioned upstream of Jλ segments, which are upstream of Cλ regions. This nested arrangement allows multiple diverse segments to be contained within a single functional genetic unit that operates coherently.
Solution Approach 2:
The patent changes the genetic composition parameters of the immunoglobulin locus by incorporating multiple human Vλ gene segments (increasing the number from typical single or few segments to multiple), human Jλ segments, and human Cλ region genes, thereby transforming the locus into a multi-component humanized structure that maximizes repertoire diversity.
3Reliability
If human Vλ, Jλ, and Cλ gene segments are operably linked, then functional human-like antibodies are produced, but the engineering process becomes more complex
Solution Approach 1:
The patent performs preliminary genetic engineering to establish the complete operable linkage of human Vλ, Jλ, and Cλ gene segments in the correct configuration before introducing the system into the rodent host. This preliminary action ensures that all necessary human genetic components are properly assembled and positioned upstream of functional rodent Cλ region genes, facilitating reliable subsequent antibody expression without requiring additional complex engineering steps.
Data Source
AI summary
Non-human animals (and/or non-human cells) and methods of using and making 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 human Igλ light chains, in whole or in part. In particular, non-human animals provided herein are, in some embodiments, characterized by expression of antibodies that contain human Igλ light chains, in whole or in part, that are encoded by human Igλ light chain-encoding sequences inserted into an endogenous Igλ light chain locus of said non-human animals. Methods for producing antibodies from non-human animals are also provided.