Transgenic Mice Common Light Chain Bispecific Antibody Production
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Solution Overview
Problem
Current methods for producing bispecific antibodies face challenges in selecting a suitable light chain that can satisfactorily associate with diverse heavy chains, relying on in vitro approaches that impose a priori restraints and risk immunogenicity, necessitating a more biologically sensible solution.
Innovation Solution
Genetically modified mice expressing human immunoglobulin heavy and light chain variable domains with a limited light chain repertoire, allowing for the production of antibodies with a common human light chain that can pair with diverse affinity-matured human heavy chains, thereby facilitating the generation of bispecific antibodies.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
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 increases complexity of selection procedures
Solution Approach 1:
The patent employs transgenic mice that autonomously perform light chain selection through their natural immune system processes. The mice are genetically engineered to express human heavy chains and a limited repertoire of human light chains, allowing in vivo self-assembly and selection of compatible pairs without external intervention or complex in vitro screening procedures
Solution Approach 2:
The patent introduces transgenic mice as an intermediary biological system between the researcher and the light chain selection process. The mice serve as living platforms that naturally facilitate the pairing of human heavy chains with compatible human light chains through their germline recombination and immune response mechanisms, eliminating the need for direct in vitro manipulation
2Adaptability or versatility
If in vitro selection methods are used to identify suitable light chains, then pairing can be achieved, but the risk of immunogenicity increases due to manipulations and modifications required
Solution Approach 1:
The transgenic mice naturally express human light chains through their engineered germline sequences, performing self-selection of compatible pairs without requiring external manipulation or modification of the light chain sequences, thereby minimizing immunogenicity risk
Solution Approach 2:
The patent pre-configures the transgenic mice with a limited repertoire of human light chain sequences in their germline during organism development, allowing natural selection and affinity maturation to occur in vivo before the actual antibody production, which reduces the need for subsequent in vitro modifications
3Productivity
If a limited light chain repertoire is used in transgenic mice, then production of antibodies with common light chain is enabled, but the scope of available light chains is restricted
Solution Approach 1:
The patent designs the transgenic mice to express a limited set of human light chains that can universally pair with multiple different human heavy chains through natural compatibility, allowing a single light chain repertoire to serve multiple antibody specificities and enabling efficient production of various bispecific antibodies
Solution Approach 2:
The patent optimizes the light chain repertoire by selecting specific human light chain sequences with high pairing compatibility across diverse heavy chains, changing the parameter from broad diversity to targeted compatibility, which enhances productivity while maintaining sufficient versatility for bispecific antibody generation
Data Source
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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 express just one or a few immunoglobulin light chain variable domains from a limited repertoire in their germline. Methods for making light chain variable regions in mice, 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, are provided.