Complementome Assay Using GluC Digestion for Homologous Protein Detection
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Solution Overview
Problem
Current methods struggle to accurately measure the levels of complement proteins such as FHR1, FHR2, FHR3, FHR4, FHR5, FH, and FHL-1 in human plasma due to their high sequence homology, which hinders the diagnosis and treatment of complement-related disorders like macular degeneration.
Innovation Solution
A method using mass spectrometry with endoproteinase GluC digestion to produce distinct peptides for each protein, allowing for the simultaneous detection and measurement of these proteins, including their breakdown products, enabling accurate diagnosis and treatment of complement-related disorders.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional methods are used to measure complement proteins, then the measurement process is simple, but the measurement precision is poor due to high sequence homology
Solution Approach 1:
The patent applies segmentation by digesting complement proteins into smaller peptides using endoproteinase GluC. This peptide-level analysis allows differentiation of proteins with high sequence homology (FHR1-FHR5, FH, FHL-1) that cannot be distinguished at the whole-protein level. Each protein produces a unique peptide fingerprint pattern that enables precise identification and quantification.
Solution Approach 2:
The patent uses endoproteinase GluC as an intermediary enzyme to convert the difficult-to-distinguish complement proteins into distinguishable peptides. This intermediary step transforms the measurement problem from direct protein detection to peptide detection, where mass spectrometry can resolve the subtle differences between highly homologous sequences.
2Measurement precision
If mass spectrometry with GluC digestion is used, then the measurement precision improves, but the device complexity and method complexity increase
Solution Approach 1:
The patent applies preliminary action by performing GluC digestion of complement proteins before mass spectrometry analysis. This pre-processing step creates a standardized peptide profile for each complement protein, enabling consistent and reproducible measurements. The digestion step is performed in advance to transform complex proteins into analyzable peptides with predictable mass patterns.
Solution Approach 2:
The patent replaces conventional immunoassay methods (which rely on antibody-antigen interactions) with mass spectrometry-based detection. This substitution eliminates the need for specific antibodies against each complement protein and uses physical mass measurement instead, providing more accurate differentiation of highly homologous proteins.
3Reliability
If current methods are used, then the diagnostic process is straightforward, but the diagnostic accuracy is insufficient for complement-related disorders
Solution Approach 1:
The patent applies universality by using a single mass spectrometry method with GluC digestion to simultaneously measure multiple complement proteins (FHR1-FHR5, FH, FHL-1) and their breakdown products. This multi-functional approach replaces multiple separate assays, improving diagnostic accuracy while actually reducing total analysis time through parallel measurement of all targets in one experiment.
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 detection and quantification of complement proteins, facilitating the identification of individuals at risk for complement-related disorders and guiding targeted therapies, improving diagnostic accuracy and treatment efficacy.
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 of identifying subjects having complement-related disorders, or at risk of such disorders, are disclosed. Also disclosed are methods for selecting subjects for treatment with complement-targeted therapies, and methods of treatment of subjects with such therapies.