Triplex Scaffold Polypeptides for High-Affinity Binding

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Solution Overview

Problem

Current protein-based binding reagents, such as monoclonal antibodies, face challenges with low affinity and high mitogenic effects, which limit their therapeutic efficacy and stability, especially when targeting multiple antigens or requiring reduced immune activation.

Innovation Solution

Development of trimeric protein complexes comprising triple helix coil scaffold domains fused with heterologous binding domains, allowing for high affinity, low mitogenic effect, and multivalent or multi-specific binding capabilities, enhancing stability and therapeutic efficacy.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If conventional monoclonal antibodies are used for binding to antigens, then they can provide immunosuppressive or therapeutic effects, but they exhibit high mitogenic effects causing adverse side effects and limited therapeutic efficacy

Engineering Contradiction:
Improvetherapeutic efficacyVSAvoidmitogenic effects
Core Design Contradiction:
ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent extracts and removes the Fc region from the antibody structure, retaining only the antigen-binding Fab regions. This extraction eliminates the mitogenic effects associated with Fc region interactions while preserving the therapeutic antigen-binding capability. The resulting F(ab')2 or single-chain antibody fragments provide therapeutic efficacy without the harmful mitogenic side effects of conventional IgG antibodies.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The patent introduces triplex scaffolds as intermediary structures that replace the Fc region's function of mediating effector activities. These scaffolds serve as structural intermediaries that maintain antibody stability and solubility while preventing unwanted Fc-mediated mitogenic interactions. The triplex scaffold acts as a neutral intermediary that supports the binding domains without triggering harmful immune responses.

Inventive Principle:
Principle #24Intermediary (Mediator)

2Object-affected harmful factors

If humanization of murine mAbs is performed using CDR grafting, then immunogenicity is reduced, but binding affinity decreases compared to murine counterparts

Engineering Contradiction:
ImproveimmunogenicityVSAvoidbinding affinity
Core Design Contradiction:
Object-affected harmful factorsVSReliability

Solution Approach 1:

The patent segments the antibody into separate functional modules: the antigen-binding CDR regions are isolated and grafted onto human framework regions, while the Fc region is completely removed. This segmentation allows independent optimization of each module - the CDRs retain their high-affinity binding characteristics from murine antibodies, while the human framework reduces immunogenicity, and the removed Fc eliminates mitogenic effects.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The patent applies local quality by preserving the murine CDR regions with their high-affinity binding properties while replacing only the immunogenic framework regions with human sequences. The critical antigen-binding local regions maintain their original high-affinity characteristics, while the non-critical framework regions are humanized to reduce immunogenicity without affecting binding affinity.

Inventive Principle:
Principle #3Local quality

3Reliability

If multimerization of antigen binding sites is implemented, then overall binding strength (avidity) increases, but the complexity of antibody structure and engineering increases

Engineering Contradiction:
Improvebinding affinityVSAvoidstructure complexity
Core Design Contradiction:
ReliabilityVSDevice complexity

Solution Approach 1:

The patent merges multiple antigen-binding sites into a single polypeptide chain or complex through systematic design. By combining two Fab regions to form F(ab')2 or by designing single-chain antibodies with multiple CDRs, the patent achieves multivalent binding (increased avidity) while maintaining relatively simple structural organization based on well-understood antibody frameworks, thus balancing enhanced binding strength with manageable structural complexity.

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The patent creates universal modular antibody fragments that can be configured in multiple valencies (monovalent, bivalent, multivalent) using the same basic F(ab')2 or single-chain architecture. This universal design allows the same structural framework to provide different binding strengths and specificities by simply varying the number and arrangement of CDR regions, reducing the complexity increase that would otherwise accompany multimerization.

Inventive Principle:
Principle #6Universality (Multi-functionality)

4Adaptability or versatility

If conventional IgG antibodies are used, then they provide bivalent binding capability, but they cannot bind to more than two different antigens simultaneously

Engineering Contradiction:
Improvemulti-specific binding capabilityVSAvoiddosage requirement
Core Design Contradiction:
Adaptability or versatilityVSQuantity of substance

Solution Approach 1:

The patent designs universal polypeptide platforms that can incorporate multiple different antigen-binding CDR regions within a single molecule. These multi-specific antibodies can simultaneously bind to multiple different antigens (e.g., CD3 and tumor-associated antigens), providing versatile therapeutic functionality. This multi-functionality allows a single dosage to achieve multiple therapeutic effects, reducing the need for combination therapies and associated dosing complexity.

Inventive Principle:
Principle #6Universality (Multi-functionality)

Data Source

PatentEP1798240B1Recombinant triplex scaffold-based polypeptides
Publication Date: 2011.04.27 IND TECH RES INST
  • EP1798240B1 patent drawingFigure 1
  • EP1798240B1 patent drawingFigure 2A~2B
  • EP1798240B1 patent drawingFigure 3

AI summary

Disclose is a protein complex that includes triple-helix-coil forming fusion polypeptide chains, each having a scaffold domain and a heterologous domain. Also disclosed are related isolated fusion polypeptide, nucleic acids, vectors, host cells, and preparation methods.