Designed Ankyrin Repeat Domains for Dual-Specificity Molecular Switches
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current molecular drug switch approaches are time-, cost-, and labor-consuming, and limited in their capabilities and applications, making them inefficient for controlling protein activity in protein-based therapeutics.
Innovation Solution
The development of designed repeat domains, specifically designed ankyrin repeat domains, that have dual binding specificity for two targets and exhibit mutually exclusive binding, allowing for the creation of molecular on/off switches to control the activation or deactivation of therapeutic agents.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If conventional molecular drug switch approaches are used, then drug activation control is achieved, but the process is time-consuming, labor-intensive, and limited in application scope
Solution Approach 1:
The designed repeat domain serves multiple functions: it provides dual binding specificity for two different targets simultaneously, enables conditional drug activation through mutually exclusive binding, and can be applied across various therapeutic contexts. This multi-functionality resolves the contradiction by expanding application scope while maintaining efficient switch mechanism development.
Solution Approach 2:
The invention changes the binding parameters of the repeat domain by engineering it to exhibit mutually exclusive binding behavior between two targets. This parameter change enables the domain to function as a molecular switch, resolving the contradiction by providing a versatile control mechanism that works across different therapeutic applications without requiring separate development for each case.
2Adaptability or versatility
If dual binding specificity is engineered into repeat domains, then molecular switch capability is achieved, but the design and construction complexity increases
Solution Approach 1:
The dual-specificity repeat domain is constructed by segmenting and combining modules from two different parent repeat domains. Each parent domain contributes specific binding characteristics, and their combination creates the dual-specificity switch capability. This segmentation approach resolves the contradiction by systematically building complex functionality from simpler, well-characterized components.
Solution Approach 2:
The invention merges two parent repeat domains with different binding specificities into a single hybrid domain that exhibits mutually exclusive binding to both targets. This merging strategy resolves the contradiction by integrating the molecular switch capability directly into the repeat domain structure, achieving versatility without requiring separate complex control systems.
3Manufacturing precision
If parental repeat domains are combined to create dual-specificity domains, then binding control precision is improved, but the manufacturing process becomes more complex
Solution Approach 1:
The hybrid repeat domain is created by copying and recombining the amino acid sequences from two parental domains. This copying approach preserves the proven binding characteristics of each parent while creating a new domain with dual specificity. The method resolves the contradiction by using established sequence templates that can be efficiently produced through standard recombinant DNA techniques.
Solution Approach 2:
The invention changes the sequence parameters of the repeat domain by introducing specific mutations and combinations from parental domains. These parameter changes are designed to create mutually exclusive binding while maintaining manufacturability through standard protein expression systems. The resolved contradiction achieves precise binding control with production complexity comparable to conventional antibody engineering.
Data Source
AI summary
The present invention relates to a method of producing a designed repeat domain which has binding specificity for two targets, wherein binding of such repeat domain to its two targets is mutually exclusive. Said designed repeat domain is preferably a designed ankyrin repeat domain. The invention also provides such designed repeat domains and recombinant proteins comprising such repeat domains, as well as the use of such repeat domains as molecular switches, such as, e.g., switches to control activation or deactivation of a therapeutic agent. The invention further relates to nucleic acids encoding such repeat domains or recombinant proteins, pharmaceutical compositions comprising such recombinant proteins, repeat domains or nucleic acids, recombinant expression vectors and host cells, and the use of such proteins, nucleic acids or pharmaceutical compositions in methods for treating diseases, such as cancer.