Single-Chain Bispecific Antibodies to Prevent Chain Mispairing
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Solution Overview
Problem
Existing bispecific antibodies face issues with mispairing of antibody heavy and light chains during production, leading to non-functional byproducts, hindering the development of clinically sufficient amounts and purity of multi-specific antibodies for cancer treatment.
Innovation Solution
Development of fusion proteins where heavy and light chains of antibodies are linked through linkers, forming single-chain bispecific or trispecific antibodies that specifically bind to mesothelin, CD3, and EGFR, respectively, to address mispairing and enhance tumor cell killing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If co-expression of heavy and light chains of different specificities is performed during bispecific antibody production, then multi-specific antibody functionality is achieved, but mispairing occurs resulting in non-functional byproducts
Solution Approach 1:
The patent divides the antibody production system into separate modules: the first antibody (anti-mesothelin) and second antibody (anti-CD3) are produced as separate entities with distinct heavy and light chains. This segmentation allows independent expression and assembly, reducing mispairing while maintaining bispecific functionality through controlled heterodimer formation of the Fc regions.
Solution Approach 2:
The patent introduces the Fc region as an intermediary element that mediates the assembly of bispecific antibodies. By engineering the Fc regions to form heterodimers through disulfide bonds or non-covalent interactions, the system enables specific pairing of heavy chains from different antibodies while light chains pair accordingly, preventing random mispairing and ensuring functional bispecific antibody production.
2Reliability
If conventional bispecific antibody production methods are used, then immune cells can be targeted, but production yield and purity are insufficient for clinical use
Solution Approach 1:
The patent merges the production advantages of monoclonal antibodies with the functional benefits of bispecific antibodies. By using the well-established, high-yield production platforms for conventional monoclonal antibodies to produce the constituent antibodies, then assembling them into bispecific formats, the method achieves both high productivity and high reliability for clinical applications.
Solution Approach 2:
The patent optimizes production parameters including expression systems, purification conditions, and assembly conditions to maximize yield and purity. By carefully controlling pH, temperature, ionic strength, and other parameters during heterodimer formation and purification steps, the method achieves clinically sufficient production levels while maintaining high purity of functional bispecific antibodies.
Data Source
AI summary
A fusion protein including a fragment of an anti-mesothelin antibody, an anti-CD3 antibody or an anti-EGFR antibody; a bispecific antibody that is specific to mesothelin and CD3; a trispecific antibody that is specific to mesothelin, CD3 and EGFR are provided. The fusion protein is effective in treating cancer. The bispecific or trispecific antibody can be prepared in a high yield and with high purity, and has excellent tumor killing and growth inhibitory effects, and thus can be effectively used in cancer treatment.