Modular Recombinant Antibody Assembly via Cohesin-Dockerin
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Solution Overview
Problem
Current methods for producing fully functional recombinant monoclonal antibodies (rAbs) complexed with antigens face challenges, particularly with multiple antigen coding regions, leading to poor or null expression due to secretion issues in mammalian cell systems.
Innovation Solution
The use of cohesin-dockerin protein domains and their surrounding linker sequences allows for the controlled assembly of rAb-antigen complexes, enabling efficient secretion and production in mammalian expression systems by leveraging the high affinity and specificity of the cohesin-dockerin interaction, even when multiple antigens are involved.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If multiple antigen coding regions are included in rAb-antigen fusion proteins, then the vaccine can target multiple pathogens or antigens, but the expression and secretion efficiency decreases significantly
Solution Approach 1:
The invention divides the complex rAb-antigen fusion protein into separate components: the rAb is expressed independently, and multiple antigens are expressed separately and then assembled onto the rAb through cohesin-dockerin interactions. This segmentation allows each component to be optimized for its own expression and secretion, avoiding the secretion problems associated with large multivalent fusion proteins while maintaining the ability to target multiple antigens.
Solution Approach 2:
The invention introduces cohesin-dockerin interaction modules as intermediaries between the rAb and multiple antigens. These modular domains facilitate the assembly of separately expressed rAb and antigen components into functional multivalent complexes, enabling efficient production of each component followed by controlled assembly, thereby resolving the contradiction between multivalent capability and expression efficiency.
2Adaptability or versatility
If rAb-antigen fusion proteins are engineered with multiple antigen coding regions, then the vaccine coverage is expanded, but the secretion from mammalian cell systems fails or is severely reduced
Solution Approach 1:
The invention segments the vaccine construct into separate expressible units (rAb and individual antigens) that can each be reliably secreted from mammalian cells, eliminating the secretion failure problem associated with large multivalent fusion proteins while maintaining expanded vaccine coverage through subsequent assembly of the separate components.
Solution Approach 2:
Cohesin-dockerin modules serve as intermediaries that enable the reliable assembly of separately secreted rAb and antigen components into functional multivalent complexes, ensuring both reliable secretion of individual components and successful formation of the final multivalent vaccine product.
3Ease of operation
If cohesin-dockerin fusion proteins are produced in mammalian secretion systems, then the high affinity and specificity interaction can be leveraged for controlled assembly, but the secretion of these bacterial protein domains is not previously established
Solution Approach 1:
The invention adapts bacterial cohesin-dockerin protein domains for expression in eukaryotic mammalian cell systems by optimizing expression conditions and verifying functionality. The cohesin and dockerin domains maintain their high affinity and specificity interactions when produced in mammalian cells, enabling controlled assembly of rAb-antigen complexes while expanding the manufacturing flexibility to use mammalian secretion systems.
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 facilitates the development of novel protein engineering formats and tools for research and clinical applications, including vaccines and cancer therapy, by ensuring the successful expression and secretion of rAb-antigen complexes, overcoming previous limitations in multivalent antibody production.
Implementation Method 1
leveraging the high affinity and specificity of the cohesin-dockerin interaction
Data Source
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Figure 4B~5
AI summary
The present invention includes compositions and methods for designing, making and using modular recombinant antibodies or fragments thereof with one half of a cohesin-dockerin pair that permits the rapid assembly of multivariant antigen conjugates.