Transgenic Mouse Antibodies Using Rearrangement-Resistant Human Light Chains
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Solution Overview
Problem
Existing methods for producing antibodies, such as humanized and chimeric monoclonal antibodies, face challenges with immunogenicity when used in humans, leading to adverse reactions and reduced efficacy due to the immune system recognizing non-human Ig sequences.
Innovation Solution
The development of transgenic murine mammals with integrated rearranged human immunoglobulin light chain variable regions in a locus resistant to DNA rearrangement and somatic hypermutation, allowing these antibodies to pair with murine heavy chains, thereby reducing immunogenicity while maintaining high affinity and specificity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If humanized or chimeric monoclonal antibodies are used in humans, then therapeutic efficacy is improved, but immunogenicity increases causing adverse reactions
Solution Approach 1:
The antibody molecule is segmented into different species components: human heavy chains and murine light chains. This segmentation allows the constant regions (human) to provide therapeutic efficacy while the variable regions (murine) maintain high affinity binding, thereby reducing immunogenicity while preserving therapeutic function
Solution Approach 2:
Different parts of the antibody molecule have different species origins optimized for their specific functions: human constant regions provide structural stability and reduced immunogenicity, while murine variable regions provide high affinity antigen binding. This local quality differentiation resolves the contradiction between efficacy and immunogenicity
2Object-affected harmful factors
If transgenic mice with integrated human light chain genes are used, then immunogenicity is reduced, but the complexity of the production system increases
Solution Approach 1:
The human light chain variable region genes are pre-integrated into the murine genome at specific loci (such as the ROSA26 locus) before immunization. This preliminary genetic modification ensures that the transgenic mice naturally produce hybrid antibodies with reduced immunogenicity, eliminating the need for post-production humanization procedures
Solution Approach 2:
The transgenic mouse serves as a living bioreactor and intermediary system that naturally produces the desired hybrid antibodies. This intermediary organism simplifies the overall production system by combining genetic engineering with natural antibody production mechanisms, avoiding complex in vitro humanization processes
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach enables the production of high-affinity, specific antibodies with reduced immunogenicity by using transgenic mice that express human light chains capable of pairing with murine heavy chains, suitable for human therapeutic applications.
Implementation Method 1
B cells are able to introduce mutations into the antibody V regions that they express, a process called somatic hypermutation. These activated B cells can also now target a somatic mutation process to their rearranged antibody gene segments and thus allow the production of daughter cells which make variants of the antibodies of the primary response.
Implementation Method 2
The genes that encode H and L chain V regions are assembled somatically from segments of germline DNA during precursor B (pre-B) cell differentiation: V, D and J gene segments for the H chain and V and J gene segments for the L chain.
Data Source
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AI summary
The invention provides transgenic, non-human animals comprising a nucleic acid encoding an immunoglobulin light chain, whereby the immunoglobulin light chain is human, human-like, or humanized. The nucleic acid is provided with a means that renders it resistant to DNA rearrangements and/or somatic hypermutations. In one embodiment, the nucleic acid comprises an expression cassette for the expression of a desired molecule in cells during a certain stage of development in cells developing into mature B cells. The invention further provides methods for producing an immunoglobulin from the transgenic, non-human animal.