Bispecific Antibody Linker-Hinge Domain Design
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Solution Overview
Problem
Existing methods for cross-linking biological molecules often fail to preserve their biological activities, leading to instability and reduced effectiveness in combined molecules.
Innovation Solution
The use of novel linker-hinge domains (LHDs) with specific sequences for cross-linking biomolecules, allowing for disulfide bridge formations and maintaining biological activities, stability, and pharmacokinetic properties in bispecific or multi-specific antibodies.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If chemical linking agents are used to cross-link biomolecules, then the molecules can be combined to form bispecific antibodies, but the biological activities of the combined molecules are not preserved
Solution Approach 1:
The patent introduces a peptide linker sequence (GGGGS) as an intermediary component that mediates the connection between biomolecules. This linker serves as a bridge that allows cross-linking while maintaining the biological activities of the combined molecules, resolving the contradiction between ease of manufacture and reliability of biological function.
Solution Approach 2:
The patent changes the chemical parameters of the cross-linking agent by using a peptide-based linker with specific amino acid sequences instead of traditional chemical linking agents. This parameter change enables the cross-linking process to preserve biological activities while maintaining ease of manufacture.
2Ease of manufacture
If traditional cross-linking methods are used, then biomolecules can be combined, but the combined molecules exhibit reduced stability
Solution Approach 1:
The patent creates a composite structure by combining the peptide linker sequence (GGGGS) with hinge sequences containing cysteine residues. This composite design provides both the flexibility needed for molecule combination and the disulfide bond formation required for enhanced stability of the bispecific antibodies.
3Ease of manufacture
If chemical cross-linking is performed, then bisspecific antibodies can be produced, but production yields are reduced
Solution Approach 1:
The patent enables self-service cross-linking by incorporating cysteine residues in the hinge sequences that can spontaneously form disulfide bonds under physiological conditions. This eliminates the need for complex external cross-linking reagents and processes, thereby improving production yields while maintaining ease of manufacture.
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
The LHDs enhance the production yields, stability, and biological functionalities of bispecific antibodies, including improved antigen-binding capabilities and cytotoxicity against tumor cells, while maintaining solubility and pharmacokinetic advantages.
Implementation Method 1
comprises cysteine-proline-proline-cysteine-proline (CPPCP; SEQ ID NO: 8)... The biomolecule maintains a T-lymphocyte activation capability, or the biomolecule maintains an antibody to antigen binding capability... comprises a dimer of the above-described protein having disulfide linkages between the hinge sequences of the dimer
Data Source
AI summary
This invention relates to bispecific antibodies having combinations of linker and hinge sequences to create linker-hinge interface domains with biological significance. Such linker-hinge interface domains covalently join two molecules, maintain the biological activities of linked molecules (target binding), stabilize the biological characteristics of new molecule (solubility and 4° C. stability), maintain the chemical, biochemical and physical properties (cytotoxicity) of the linked molecules, and modulate the biological characteristics of the linked molecules (activating T-lymphocytes without significant sign of proliferations). Both linker (GGGGS) and hinge (CPPCP) sequences are required to establish functional linker-hinge interface domains as deletion of any of the component resulted in significant lost of T-lymphocyte mediated activity.