Transgenic Animal Humanized Antibody Expression
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Solution Overview
Problem
Current methods for producing humanized antibodies in non-human transgenic animals face challenges such as low expression levels due to species-specific regulatory elements and interference from endogenous antibody production, which limits the yield and effectiveness of human immunoglobulin production.
Innovation Solution
The use of transgene constructs encoding chimeric Igα and Igβ subunits with high sequence identity to human counterparts, combined with a humanized immunoglobulin locus, to promote normal B-cell development and reduce endogenous antibody production, thereby enhancing the expression and yield of humanized antibodies in non-human animals like rabbits and chickens.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If transgenes encoding humanized immunoglobulin loci are introduced into non-human animals, then humanized antibody production is achieved, but expression levels remain low due to species-specific regulatory elements
Solution Approach 1:
The patent modifies regulatory parameters by replacing species-specific regulatory elements with human homologs. Specifically, it introduces human Igα and Igβ transgenes that contain human promoter regions, enhancers, and other regulatory sequences, thereby changing the regulatory parameters from non-human to human-type control mechanisms. This enables the humanized immunoglobulin loci to be expressed at higher levels in non-human transgenic animals by providing appropriate human regulatory element recognition sequences.
2Quantity of substance
If endogenous antibody production is suppressed to enhance human immunoglobulin expression, then humanized antibody yield increases, but B-cell development and survival are compromised
Solution Approach 1:
The patent segments the B-cell receptor complex into distinct transgenic components: humanized immunoglobulin loci (heavy and light chains) combined with separate transgenes encoding Igα and Igβ. This segmentation allows independent optimization of each component - the humanized immunoglobulins provide the antigen-binding specificity while the human Igα/Igβ provide the necessary signaling functions for B-cell development and survival. By dividing the system into functional modules, the patent achieves both high humanized antibody yield and maintained B-cell viability.
Solution Approach 2:
The patent introduces human Igα and Igβ as intermediary components that mediate between the humanized immunoglobulin loci and the non-human animal's cellular machinery. These intermediary proteins serve as adapters that enable proper signaling through the B-cell receptor complex, facilitating both B-cell development/survival and high-level expression of humanized antibodies. The human Igα/Igβ act as essential mediators that bridge the gap between humanized antibody components and the host animal's immune system.
Data Source
AI summary
The present invention describes transgenic animals with human(ized) immunoglobulin loci and transgenes encoding human(ized) Igα and/or Igβ sequences. Of particular interest are animals with transgenic heavy and light chain immunoglobulin loci capable of producing a diversified human(ized) antibody repertoire that have their endogenous production of Ig and/or endogenous Igα and/or Igβ sequences suppressed. Simultaneous expression of human(ized) immunoglobulin and human(ized) Igα and/or Igβ results in normal B-cell development, affinity maturation and efficient expression of human(ized) antibodies.