Repeat-Chain Scaffold for Antibody-Toxin Dimer Yield
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Conventional methods for producing inter-monomer disulfide bond bridged dimers have low yields due to inefficient formation of inter-monomer disulfide bond bridges during the refolding process, limiting the production of high-affinity antibody-toxin complexes for cancer treatment and diagnostic applications.
Innovation Solution
The use of repeat-chains with repeated affinity domains binding specifically to monomers creates high local concentrations and increased collision frequencies between monomers, facilitating the formation of inter-monomer disulfide bond bridges within repeat-chain/multiple-monomer complexes, significantly enhancing the yield of disulfide bond bridged dimers.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional refolding methods are used to produce inter-monomer disulfide bond bridged dimers, then the production process is simple, but the yield is low due to inefficient formation of inter-monomer disulfide bond bridges
Solution Approach 1:
The patent introduces a repeat-chain containing multiple affinity domains as an intermediary binding matrix. This repeat-chain mediates the association of multiple monomers by providing multiple binding sites, thereby facilitating the formation of inter-monomer disulfide bond bridges. The repeat-chain acts as a scaffold that brings monomers into close proximity, increasing the efficiency of dimer formation from less than 1% to over 20%.
Solution Approach 2:
The patent segments the binding function into multiple independent affinity domains arranged in a repeat-chain structure. Each affinity domain can independently bind to a monomer, allowing the system to accommodate multiple monomers simultaneously. This segmentation of binding sites enables controlled multimerization and significantly improves dimer yield compared to conventional refolding methods.
2Productivity
If repeat-chains with multiple affinity domains are used as binding matrix, then the yield of dimers is significantly increased, but the complexity of the system increases
Solution Approach 1:
The repeat-chain is designed with multiple identical affinity domains that can universally bind to the same type of monomer. This multi-functionality allows a single repeat-chain structure to accommodate multiple monomers, enabling scalable production of dimers and higher-order multimers. The universal binding capability simplifies the manufacturing process by using a standardized repeat-chain design for various applications.
3Productivity
If monomers are allowed to freely diffuse during refolding, then the refolding process is simple, but the collision frequency between monomers is low resulting in low dimer formation efficiency
Solution Approach 1:
The patent applies preliminary action by pre-organizing monomers on the repeat-chain binding matrix before dimer formation occurs. The repeat-chain holds multiple monomers in close proximity in a pre-assembled complex, ensuring that when refolding conditions are applied, the monomers are already positioned for efficient disulfide bond formation. This preliminary organization dramatically increases collision frequency compared to free diffusion.
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 method achieves a 200-fold increase in the yield of disulfide bond bridged dimers, resulting in antibody-toxin complexes with higher antigen binding strength, cytotoxicity, and stability, suitable for cancer treatment and improved diagnostic sensitivity.
Implementation Method 1
repeat-chains with repeated affinity domains binding specifically to monomers creates high local concentrations and increased collision frequencies between monomers
Implementation Method 2
formation of inter-monomer disulfide bond bridges within repeat-chain/multiple-monomer complexes
Data Source
AI summary
The present invention relates to a method for manufacturing multimers by making repeat-chains comprising repeatedly linked affinity domains binding specifically to monomers, and by using the same to create a repeat-chain/multiple-monomer complex created from the repeat-chains and a multiple number of monomers, thereby facilitating the formation of bond bridges between the monomers in the complex to produce inter-monomeric bond bridged multimer.The present invention relates to a super-complex prepared by cross-binding between repeat-chain/multiple-monomer complexes, and a method for amplifying the effect of monomer through the formation of the said super-complex. Particularly, the repeat-chain/multiple-monomer complex is prepared by containing repeat-chains of binding domain having binding specificity to monomers as active ingredients, and then the super-complex is prepared by cross-binding between such complexes.