Marker Peptide TLCAGVMEGGIDTCNR for SVTLEs Detection
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Solution Overview
Problem
Current methods for characterizing and quantifying snake venom thrombin-like enzymes (SVTLEs) from Agkistrodon Halys Pallas are complex and inaccurate, lacking a reliable method for determining the source and content, which is crucial for quality control and clinical effectiveness.
Innovation Solution
A marker peptide with the amino acid sequence TLCAGVMEGGIDTCNR is identified and used in a liquid chromatography-mass spectrometry method involving trypsin enzymolysis and multiple-reaction monitoring to accurately detect and quantify SVTLEs, enabling the determination of the source and content of SVTLEs in samples.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If pulsed field gel electrophoresis or efficacy testing method is used to characterize SVTLEs content, then the detection can be performed, but the method is complicated and poor in accuracy
Solution Approach 1:
The patent extracts a specific marker peptide (TLCAGVMEGGIDTCNR) from the complex SVTLEs protein structure. By detecting only this specific peptide fragment through LC-MS/MS, the method simplifies the analysis from characterizing entire complex proteins to detecting a single specific peptide sequence, thereby reducing method complexity while improving measurement precision
Solution Approach 2:
The patent changes the detection parameter from analyzing entire proteins (complex, low precision) to analyzing a specific peptide fragment with unique amino acid sequence (simple, high precision). The marker peptide's specific sequence TLCAGVMEGGIDTCNR serves as a unique identifier that transforms the detection approach from complex protein characterization to precise peptide quantification
2Measurement precision
If current quality standard methods are used, then the source of species can be detected, but there is no specific method for detecting thrombin-like enzyme content
Solution Approach 1:
The patent applies local quality by focusing detection on a specific local region (marker peptide TLCAGVMEGGIDTCNR) within the SVTLEs protein rather than analyzing the entire protein. This localized approach provides high specificity for thrombin-like enzyme detection while simplifying the measurement process through targeted peptide quantification
Solution Approach 2:
The marker peptide serves as an intermediary between the complex SVTLEs protein and the detection system. By detecting this specific peptide fragment that is unique to Agkistrodon Halys Pallas SVTLEs, the method indirectly quantifies the parent protein with high precision, making the detection process easier while maintaining high specificity
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 provides a simple, rapid, and accurate means for quality control of Agkistrodon Halys Pallas venom, enhancing the reliability and safety of clinical SVTLE products by distinguishing the source and quantifying the thrombin-like enzyme content.
Implementation Method 1
pretreating the to-be-detected sample by trypsin through enzymolysis
Implementation Method 2
liquid chromatography-mass spectrometry method
Implementation Method 3
liquid chromatography-mass spectrometry method involving trypsin enzymolysis and multiple-reaction monitoring
Data Source
AI summary
The present invention relates to the field of chemical analysis detection and application, in particular to a marker peptide of snake venom thrombin-like enzymes (SVTLEs) from Agkistrodon Halys Pallas and an application thereof. The amino acid sequence of the marker peptide of snake venom thrombin-like enzymes (SVTLEs) from Agkistrodon Halys Pallas is TLCAGVMEGGIDTCNR. Characterizing the source of species and a content of the SVTLEs in a to-be-detected sample by using the marker peptide includes the following steps of: pretreating the to-be-detected sample by trypsin through enzymolysis, and taking a supernatant of an enzymolysis liquid as a test solution; and injecting the test solution and a reference solution into a liquid chromatography-mass spectrometer, and selecting a qualitative ion pair and a quantitative ion pair for detecting the source of species and a content of the SVTLEs in the to-be-detected sample.