Multivalent Antigen-Binding Fv Molecule Domain Order
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Solution Overview
Problem
Current tandem diabodies with the domain order V H A-V L B-V H B-V L A have limitations in biological activity and therapeutic efficacy, necessitating an improved antigen-binding molecule with enhanced cytotoxicity and reduced dosages for immunotherapy applications.
Innovation Solution
A dimeric antigen-binding molecule with the domain order V L A-V H B-V L B-V H A is developed, comprising two polypeptide chains with specific variable domains arranged to form tetravalent, bispecific antigen-binding sites, enhancing biological activity and cytotoxicity by modified crosslinking and antigen binding efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If the domain order V H A-V L B-V H B-V L A is used in tandem diabodies, then the molecule can be constructed with standard intermolecular pairing, but the biological activity and cytotoxicity are limited
Solution Approach 1:
The patent inverts the conventional domain arrangement by placing the light chain variable domain (V L A) at the N-terminus followed by the heavy chain variable domain (V H B), then light chain (V L B) and heavy chain (V H A) at the C-terminus. This reversed sequence arrangement (V L A-V H B-V L B-V H A) fundamentally changes the molecular architecture to achieve superior biological activity and cytotoxicity compared to the standard V H A-V L B-V H B-V L A configuration.
2Productivity
If the conventional V H A-V L B-V H B-V L A domain order is used, then the tandem diabody can be produced with standard protocols, but higher dosages are required for therapeutic efficacy
Solution Approach 1:
The patent changes the fundamental parameter of domain sequence arrangement from the conventional V H A-V L B-V H B-V L A to the inverted V L A-V H B-V L B-V H A. This parameter change in the molecular architecture results in enhanced cytotoxic activity and therapeutic efficacy, allowing for reduced dosages while maintaining or improving treatment effectiveness.
3Reliability
If standard tandem diabody construction is used, then the molecule has four antigen-binding sites, but the crosslinking efficiency and antigen binding capability are insufficient
Solution Approach 1:
The patent introduces asymmetry in the domain arrangement by alternating light chain and heavy chain domains in a specific sequence (V L A-V H B-V L B-V H A). This asymmetric arrangement creates optimized crosslinking patterns that enhance antigen binding efficiency and molecular stability, improving the functional performance of the tetravalent structure.
Data Source
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AI summary
In one aspect, the present invention relates to an antigen-binding molecule comprising two polypeptide chains, each polypeptide chain having at least four variable domains in an orientation preventing Fv formation and the two polypeptide chains are dimerized with one another thereby forming a multivalent antigen-binding molecule. On each of the two polypeptide chains the four variable domains are arranged in the order V L A-V H B-V L B-V H A from the N-terminal to the C-terminal of the polypeptide.. Compositions of the antigen-binding molecule and the methods of using the antigen-binding molecule or the compositions thereof for treatment of various diseases are also provided herein.