Fibronectin Conformation Antibodies for Integrin Switch Detection
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Solution Overview
Problem
There is a need for compositions and methods to detect and regulate the fibronectin-integrin switch, which is critical for diagnosing, distinguishing, treating, and preventing diseases and disorders associated with this pathway, as the relevance of mechanical forces on integrin binding profiles in vivo remains undefined.
Innovation Solution
Development of isolated and purified antibodies and scFv peptides that target specific conformational states of fibronectin, particularly the FnIII9-4G-10 state, to detect and regulate fibronectin-integrin interactions, allowing for the detection of force-induced conformational changes and treatment of associated diseases.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If mechanical forces are applied to fibronectin to induce conformational changes, then integrin binding affinity is modulated, but the structural stability of fibronectin is compromised
Solution Approach 1:
The patent employs dynamic conformational switching of fibronectin type III repeats in response to mechanical forces. The FnIII9-10 domain transitions between folded and unfolded states under force, dynamically modulating integrin binding affinity without permanent structural damage. This dynamic behavior allows the protein to adapt its binding properties based on mechanical conditions while maintaining overall structural integrity.
Solution Approach 2:
The patent utilizes changes in physical parameters (mechanical force, conformational state) to regulate integrin binding. By monitoring and responding to force-induced conformational changes in the FnIII9-10 domain, the system modulates binding affinity through parameter changes rather than permanent structural alterations, resolving the contradiction between reliability and stability.
2Measurement precision
If antibodies target specific conformational states of fibronectin to detect force-induced changes, then diagnostic precision is improved, but detection complexity increases
Solution Approach 1:
The patent employs antibodies with specificity for local conformational states of fibronectin (e.g., folded vs. unfolded FnIII9-10). Each antibody recognizes a specific local structural feature, allowing precise detection of force-induced conformational changes. This local quality approach enables high diagnostic precision by targeting specific conformational signatures without requiring complex detection systems.
Solution Approach 2:
The patent utilizes conformation-specific antibodies that can be detected through various signaling methods (e.g., fluorescent labels, enzymatic reactions). The presence of specific conformational states is indicated by the binding of these antibodies, which can be visualized through colorimetric, fluorescent, or other optical changes, simplifying the detection process while maintaining high precision.
3Adaptability or versatility
If type III repeats are made sensitive to unfolding to enable mechanosensing, then mechanosensitivity is enhanced, but structural stability deteriorates
Solution Approach 1:
The patent designs type III repeats with controlled dynamic stability, allowing them to unfold and refold in response to mechanical forces. The repeats are engineered to be stable under normal conditions but capable of reversible unfolding when subjected to physiological forces, enabling mechanosensing while maintaining overall structural integrity through dynamic equilibrium.
Solution Approach 2:
The patent utilizes periodic folding and unfolding of type III repeats in response to cyclic mechanical loading. The repeats exhibit reversible conformational transitions that occur periodically with mechanical stress cycles, allowing the system to sense mechanical forces through repeated structural transitions without permanent damage, thus enhancing mechanosensitivity while preserving stability.
Data Source
AI summary
Provided are antibodies that include amino acid sequences of SEQ ID NOs: 2, 4, and 6-12, or amino acid sequences that are about 95% identical thereto, and fragments thereof. Also provided are scFv peptides that include a VH segment having a first amino acid sequence of amino acids 4-113 of any one of SEQ ID NOs: 2 and 8-12, a VL segment having a second amino acid sequence having amino acids 113-237 of SEQ ID NOs. 2 and 8-12, or both; nucleic acids encoding the same; methods for using the same to detect and/or target conformational states of FN in samples; methods for treating diseases and/or disorders and/or for meliorating at least one symptom of consequence of a disease or disorder associated with abnormal expression of a force-induced conformational state of FN in subjects; and methods for screening for compounds having selective binding activities for conformational states of FN.