Limited-Repertoire Mice for Common-Light-Chain Bispecific Antibodies
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Solution Overview
Problem
Existing methods for creating bispecific antibodies rely on in vitro manipulations that impose a priori restraints, making it difficult to develop a suitable light chain component that can associate with different heavy chains effectively.
Innovation Solution
Genetically modified mice expressing a limited repertoire of human immunoglobulin light chain variable domains, allowing for the in vivo selection of light chains that can pair with diverse affinity-matured human heavy chains, thereby facilitating the production of multispecific antigen-binding proteins.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If in vitro methods are used to select light chains by surveying usage statistics or testing in phage display libraries, then a light chain can be identified that associates with heavy chains, but the process imposes a priori restraints and requires extensive in vitro manipulation
Solution Approach 1:
The patent employs living animal systems (mice) to autonomously perform light chain selection through natural immune responses. The mice's B cells naturally pair light chains with heavy chains under physiological conditions, eliminating the need for complex in vitro screening and manipulation. The biological system self-selects compatible light chain-heavy chain pairs through in vivo processes.
Solution Approach 2:
The patent introduces living animal organisms as an intermediary system between the researcher and the light chain selection process. The mice serve as a biological mediator that naturally performs the complex task of selecting and pairing light chains with diverse heavy chains, replacing direct in vitro manipulation with an indirect biological approach.
2Manufacturing precision
If in vitro methods with pre-selected heavy chains are used, then light chain candidates can be tested and modified, but the approach requires extensive manipulation and imposes sequence identity restraints
Solution Approach 1:
The biological system in living mice automatically optimizes light chain sequences through natural somatic hypermutation and selection processes during immune responses. The mice's immune system self-performs the complex optimization task that would otherwise require extensive in vitro sequence manipulation and testing.
Solution Approach 2:
The patent allows light chain sequences to undergo natural somatic hypermutation in vivo, changing sequence parameters through biological mutation and selection processes. This natural parameter evolution occurs within the living organism without requiring direct human manipulation of sequence parameters in the laboratory.
3Adaptability or versatility
If diverse light chain repertoires are used in conventional mice, then broad antibody specificity can be achieved, but it becomes difficult to identify and isolate specific light chains for bisspecific antibody production
Solution Approach 1:
The patent segments the light chain repertoire by engineering mice to express only a limited, defined set of human light chain variable domains alongside diverse heavy chains. This segmentation creates distinct, traceable light chain populations that can be easily identified and isolated, while still providing sufficient diversity for broad antibody specificity through the heavy chain variations.
Data Source
AI summary
A genetically modified mouse is provided, wherein the mouse expresses an immunoglobulin light chain repertoire characterized by a limited number of light chain variable domains. Mice are provided that present a choice of two human light chain variable gene segments such that the immunoglobulin light chains expresses by the mouse comprise one of the two human light chain variable gene segments. Methods for making bispecific antibodies having universal light chains using mice as described herein, including human light chain variable regions, are provided. Methods for making human variable regions suitable for use in multispecific binding proteins, e.g., bispecific antibodies, and host cells are provided.