Complement Protein Detection via GluC Digestion
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Solution Overview
Problem
Current methods struggle to accurately measure the levels of complement proteins, particularly FH, FHL-1, and FHR proteins, due to their high sequence homology, which hinders effective diagnosis and treatment of complement-related disorders such as age-related macular degeneration.
Innovation Solution
A method using mass spectrometry with endoproteinase GluC digestion to produce distinct peptides for simultaneous detection and measurement of FH, FHL-1, FHR proteins, and C3 breakdown products, enabling accurate quantification of key complement proteins in plasma.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional methods are used to measure complement proteins, then the measurement process is simpler, but the measurement precision is poor due to high sequence homology between FH, FHL-1, and FHR proteins
Solution Approach 1:
The patent segments the complement proteins into distinct peptide fragments through targeted proteolytic cleavage. By using specific proteases to cleave at unique sites within each protein, the method generates peptide segments with distinct mass spectra that can be individually identified and quantified, thereby resolving the measurement precision issue caused by high sequence homology
Solution Approach 2:
The patent changes the physical-chemical parameters of the complement proteins by converting them from intact protein form to peptide fragment form through controlled proteolysis. This parameter change (from whole protein to fragments) enables mass spectrometry to distinguish between highly similar proteins based on their unique peptide mass patterns, thereby improving measurement precision
2Measurement precision
If mass spectrometry with GluC digestion is used to distinguish between highly similar proteins, then the measurement precision improves, but the method complexity increases
Solution Approach 1:
The patent applies preliminary action by performing targeted proteolytic digestion with GluC before mass spectrometry analysis. This pre-processing step cleaves the complement proteins at specific glutamic acid residues to generate characteristic peptide patterns, making the subsequent mass spectrometry measurement more straightforward and interpretable
Solution Approach 2:
The patent introduces GluC protease as an intermediary agent that mediates the conversion of complement proteins into detectable peptide forms. This intermediary enzyme facilitates the distinction between highly similar proteins by generating unique peptide fingerprints, thereby enabling accurate measurement while maintaining procedural simplicity
3Difficulty of detecting and measuring
If conventional assays are used, then the ease of operation is higher, but the ability to distinguish between highly similar proteins is poor
Solution Approach 1:
The patent replaces conventional mechanical/chemical assay methods with mass spectrometry-based detection. This substitution enables precise distinction between highly similar complement proteins by measuring their mass-to-charge ratios and peptide fragmentation patterns, thereby reducing the difficulty of detection and measurement despite requiring more sophisticated instrumentation
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 allows for precise measurement of complement proteins and their breakdown products, facilitating the diagnosis and treatment of complement-related disorders by distinguishing between highly similar proteins and tracking protein activation and progression.
Implementation Method 1
digesting the protein(s) with endoproteinase GluC to obtain one or more peptides
Implementation Method 2
detecting the one or more peptides by mass spectrometry
Data Source
AI summary
Methods for detecting and determining the level of complement proteins are disclosed, in particular using mass spectrometry. Also disclosed are methods of identifying subjects having or at risk of developing a complement-related disorder, and methods of treating the same.