Chimeric Antibody Loci Construction in Transgenic Mammals
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Solution Overview
Problem
Existing methods for generating mice with human immune systems to produce fully human antibodies face limitations such as large insert sizes, low efficiency, and restricted repertoire of V-regions, leading to suboptimal antibody affinities and complex breeding processes.
Innovation Solution
A method for constructing chimeric human heavy and light chain loci in non-human mammals, allowing for the production of a repertoire of chimeric antibodies with human variable regions and non-human constant regions, using techniques like homologous recombination to insert human IgH V, D, and J regions upstream of the host constant region, enabling efficient production of chimeric antibodies.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If the entire heavy and light chain loci are introduced into transgenic models, then a complete human antibody repertoire can be generated, but the large insert size makes it impossible to introduce the entire loci
Solution Approach 1:
The patent divides the heavy and light chain loci into separate transgenic constructs, each containing specific V, D, and J regions. This segmentation allows the large loci to be introduced as manageable fragments while still enabling complete antibody repertoire generation through combinatorial V(D)J recombination in transgenic animals.
Solution Approach 2:
The patent embeds human V, D, and J region genes within transgenic expression constructs that include necessary regulatory elements. These nested genetic elements are integrated into the host animal genome to create functional transgenic loci that mimic the structure and function of native human immunoglobulin genes.
2Adaptability or versatility
If large insert transgenic lines are generated, then complete human antibody loci can be introduced, but the efficiency is very low and the process is complex and time-consuming
Solution Approach 1:
The patent segments the immunoglobulin loci into smaller, modular transgenic constructs that can be more efficiently introduced and integrated into the host genome. This segmentation reduces the complexity of generating transgenic lines while maintaining the ability to generate complete antibody repertoires.
Solution Approach 2:
The patent prepares transgenic constructs with all necessary regulatory elements, V, D, andJ regions pre-assembled in appropriate configurations before introduction into host animals. This preliminary structuring of transgenic loci streamlines the generation process and reduces the time required to obtain functional transgenic lines.
3Ease of manufacture
If transgenic lines are generated with limited V-region repertoires, then the transgenic models can be created, but individual antibody affinities rarely reach those obtained from intact animals
Solution Approach 1:
The patent ensures that each transgenic construct contains the appropriate local genetic elements (V, D, andJ regions with associated enhancers and regulatory sequences) necessary for generating high-affinity antibodies. This local quality preservation in transgenic loci enables the generation of antibodies with affinities comparable to those from intact human immune systems.
Solution Approach 2:
The patent optimizes various parameters of the transgenic constructs, including the arrangement of V, D, andJ regions, the inclusion of appropriate enhancer elements, and the configuration of constant regions, to enable the generation of high-affinity antibodies with diverse specificities that match or exceed the capabilities of intact animal models.
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 generation of non-human mammals capable of producing antibodies with high affinity and diverse repertoires, overcoming the limitations of previous methods by maintaining non-human mammal control sequences and enhancing antibody expression levels.
Implementation Method 1
using techniques like homologous recombination to insert human IgH V, D, and J regions upstream of the host constant region
Data Source
AI summary
The invention discloses methods for the generation of chimaeric human—non-human antibodies and chimaeric antibody chains, antibodies and antibody chains so produced, and derivatives thereof including fully humanised antibodies; compositions comprising the antibodies, antibody chains and derivatives, as well as cells, non-human mammals and vectors, suitable for use in the methods.