Prion Detection via Plasmin Cleavage of Normal Forms
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Solution Overview
Problem
Current methods for detecting pathological prions lack sensitivity, are time-consuming, and require proteinase K treatment, making early diagnosis in living animals or humans challenging.
Innovation Solution
A method involving capture antibodies that recognize both pathological and non-pathological prion proteins, followed by plasmin treatment to distinguish between the two forms, allowing for selective detection of uncleaved pathological prions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If western blot or other conventional detection methods are used, then pathological prions can be detected, but the test duration is long (7-8 hours) and requires multiple treatment steps
Solution Approach 1:
The invention extracts and utilizes the specific conformational property of pathological prions that makes them resistant to plasmin degradation. By removing the need for proteinase K treatment and other complex preparation steps, the method directly detects pathological prions through their unique resistance to plasmin, significantly reducing test duration while maintaining high detection sensitivity
Solution Approach 2:
The invention changes the detection parameter from general protein detection to conformation-specific detection using plasmin resistance. By measuring the differential degradation resistance between pathological and normal prion proteins, the method achieves rapid and specific detection without lengthy preparation protocols
2Measurement precision
If proteinase K treatment is used to detect pathological prions, then detection can be performed, but the method is complex and time-consuming
Solution Approach 1:
The invention replaces proteinase K treatment with plasmin-based detection, changing the biochemical parameter from general proteolytic resistance to specific conformation-dependent plasmin resistance. This simplifies the method by using a single enzyme with well-defined specificity rather than multiple treatment steps
Solution Approach 2:
The invention uses plasmin as an intermediary substance to differentiate pathological from normal prions. Instead of directly detecting prion conformation, the method employs plasmin as a mediator that selectively degrades normal prions, leaving pathological prions intact for detection
3Measurement precision
If enrichment methods like PMCA are used, then detection sensitivity can be improved, but the procedure becomes more complex and time-consuming
Solution Approach 1:
The invention extracts the key diagnostic feature of pathological prions (plasmin resistance) and uses it directly for detection without requiring enrichment amplification. This eliminates the time-consuming PMCA cycles while maintaining high detection sensitivity through specific immunodetection of the resistant prion form
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 enables early detection of pathological prions with high sensitivity and specificity, reducing detection time and costs, and can be automated, achieving ID50 values of less than 1,000/ml.
Implementation Method 1
The serine protease plasmin (preferred cleavage site Lys-Xaa>Arg-Xaa) is one of plasminogen, a ubiquitous zymogen precursor; synthesized enzyme that plays an important role in the conversion of fibrin into soluble products (fibrinolysis) and in the proteolytic degradation of the extracellular matrix (plasmin-induced proteolysis). Recently, it has been reported that plasmin is able to cleave PrP C
Data Source
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AI summary
The aim of the invention is to create a method for detecting pathological prions which is highly sensitive, can be carried out quickly and at a low cost, and allows prions to be detected at an early stage of a disease. Said aim is achieved by the fact that immobilized capture antibodies bind the pathological and non-pathological form of the prion protein contained in a sample, whereupon the bound non-pathological form is specifically divided by means of plasmin. The undivided, pathological form of the prion protein, which is bound by the immobilized capture antibodies, can then be easily detected with the aid of detection antibodies. The inventive method is used for detecting pathological prions.