Transgenic Rodent Producing Chimeric Human IgE Antibodies
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Solution Overview
Problem
Current methods for producing human IgE antibodies are inefficient due to the rarity and short-lived nature of IgE-producing B cells, making it difficult to generate high amounts of high-affinity chimeric human IgE antibodies, and existing transgenic mice strains cannot produce significant quantities of IgE antibodies.
Innovation Solution
A transgenic laboratory rodent is developed with human immunoglobulin mu and epsilon heavy-chain constant transgenes inserted in place of the endogenous mu heavy-chain switch sequence, allowing for site-specific recombination to switch from IgM to IgE production, enabling the generation of high-affinity chimeric human IgM and IgE antibodies.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If human IgE antibodies are isolated from patients, then high-affinity IgE antibodies can be obtained, but the amount obtained is very small and limited to specific antigens
Solution Approach 1:
The patent creates transgenic mice that copy the human immunoglobulin heavy chain locus structure, enabling these animals to naturally produce human IgE antibodies with high affinity. This copying approach allows unlimited production of high-quality antibodies without relying on patient samples.
Solution Approach 2:
The patent changes the genetic parameters of the mice by introducing human immunoglobulin heavy chain constant region genes (Cμ and Cε) and variable region genes, transforming them into factories that produce human IgE antibodies. This parameter change enables both high affinity and abundant production.
2Manufacturing precision
If recombinant human IgE is produced by cloning variable regions and rearranging with constant genes, then specific IgE antibodies can be produced, but the process is long, costly and produces only one antibody molecule at a time
Solution Approach 1:
The transgenic mice possess a universal human immunoglobulin heavy chain locus that can generate diverse IgE antibodies against multiple different antigens. A single transgenic animal serves as a platform for producing various specific IgE antibodies, eliminating the need for separate cloning processes for each antibody.
Solution Approach 2:
The transgenic mice's immune systems automatically perform the functions of antibody diversification and selection when immunized with antigens. The animals self-generate the diverse repertoire of high-affinity IgE antibodies through natural immune responses, eliminating the need for manual cloning and rearrangement operations.
3Measurement precision
If existing transgenic mouse strains with human immunoglobulin loci are used, then high-affinity antibodies can be generated, but the frequency of IgE+ B cells is extremely low making detection and extraction nearly impossible
Solution Approach 1:
The patent introduces human epsilon constant region genes specifically into the transgenic mice, creating a localized human IgE production capability. This local quality enhancement allows IgE+ B cells to be detected and studied, while maintaining high antibody affinity.
Solution Approach 2:
The patent enables detection of IgE+ B cells by introducing human immunoglobulin markers that can be identified through flow cytometry and other detection methods. The transgenic mice express human IgE antibodies that can be specifically detected, making the rare IgE+ B cells visible and extractable.
4Quantity of substance
If human IgE is produced in existing transgenic mice, then some IgE antibodies can be obtained, but the amounts are not significant
Solution Approach 1:
The patent performs preliminary genetic engineering by introducing human immunoglobulin heavy chain constant region genes (Cμ and Cε) and variable region genes into the mice before immunization. This preliminary action prepares the animals to produce high amounts of IgE antibodies efficiently upon antigen exposure.
Solution Approach 2:
The transgenic mice's immune systems automatically perform the functions of antibody diversification and selection when immunized with antigens. The animals self-generate the diverse repertoire of high-affinity IgE antibodies through natural immune responses, eliminating the need for manual cloning and rearrangement operations.
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 allows for the rapid and efficient production of high-affinity chimeric human IgM and IgE antibodies, overcoming the limitations of previous methods by generating detectable levels of functional IgE B cells and enabling studies on IgE antibody functions and applications in therapy and diagnosis.
Implementation Method 1
site-specific recombination between said first and second site-specific recombination sequences
Data Source
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AI summary
The invention relates to a transgenic non-human mammal comprising human immunoglobulin mu and epsilon heavy-chain constant transgenes Cμ and Cε inserted in place of endogenous mu heavy-chain switch sequence Sμ, and its use for producing chimeric human immunoglobulin E antibodies specific for an antigen of interest.