DPP3 Activity Measurement via Specific Inhibitor Separation
Solution Overview
Problem
Current methods fail to specifically determine active DPP3 in bodily fluid samples and effectively diagnose or treat diseases related to necrotic processes, as they often measure other aminopeptidases alongside DPP3 or inhibit DPP3 activity non-specifically.
Innovation Solution
A method involving a capture-binder that specifically binds to full-length DPP3, separating it from bodily fluids, and quantifying its activity using a substrate, while minimizing inhibition, allowing for accurate measurement of DPP3 activity in bodily fluids.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If general protease inhibitors are used to inhibit DPP3 activity, then DPP3 activity is inhibited, but other aminopeptidases are also inhibited non-specifically
Solution Approach 1:
The patent uses a specific DPP3 inhibitor as an intermediary substance that selectively binds to and inhibits only DPP3 enzyme activity without affecting other aminopeptidases. This mediator enables specific measurement of DPP3 by comparing enzyme activity with and without the inhibitor present.
Solution Approach 2:
The patent changes the chemical parameters of the inhibition approach by using a specific inhibitor molecule with defined chemical structure and binding properties that selectively interact with DPP3's active site, rather than using general protease inhibitors that affect multiple enzymes.
2Quantity of substance
If current measurement methods are used to determine aminopeptidase activity, then total aminopeptidase activity is measured, but active DPP3 cannot be specifically determined
Solution Approach 1:
The patent extracts the specific DPP3 component from the total aminopeptidase mixture by using a specific inhibitor that selectively binds to DPP3. This allows the activity measurement to focus exclusively on DPP3 rather than measuring all aminopeptidases together.
Solution Approach 2:
A specific DPP3 inhibitor serves as a mediator that selectively interacts with DPP3 to enable its specific quantification among other aminopeptidases.
3Object-generated harmful factors
If non-specific protease inhibitors are used, then enzyme activity can be inhibited, but the inhibition lacks specificity for DPP3
Solution Approach 1:
The patent applies local quality by designing an inhibitor with specific chemical properties that match only the DPP3 enzyme's active site characteristics, creating a localized and selective interaction rather than broad non-specific inhibition.
Solution Approach 2:
The patent changes the inhibition parameter from general protease inhibition to specific DPP3 inhibition by selecting an inhibitor with molecular characteristics that selectively bind to DPP3's unique structural features.
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 approach enables precise determination of active DPP3 in bodily fluids, facilitating the diagnosis of diseases related to necrotic processes and providing a basis for targeted therapeutic interventions.
Implementation Method 1
Contacting said sample with a capture-binder that binds specifically to full-length DPP3
Implementation Method 2
DPP3 activity can be measured by addition of the fluorogenic substrate Arg-Arg-βNA and monitoring fluorescence in real time
Implementation Method 3
Quantifying DPP3 activity by measuring the conversion of the substrate of DPP3
Data Source
AI summary
The present invention is directed to methods for determining active DPP3 in a bodily fluid sample, an assay or kit for determining active DPP3 in a bodily fluid sample, a method for diagnosing a disease or condition in a subject accompanied by or related to necrotic processes and methods of treating or preventing said disease.