Mass Spectrometry Allergen Quantitation Using Internal Standard Peptides
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Solution Overview
Problem
Current methods for screening plant seed proteins for allergens are inefficient and lack accuracy, particularly in distinguishing between natural and genetically-modified crops, and require extensive resources such as gel electrophoresis and antibodies.
Innovation Solution
A high-throughput method utilizing mass spectrometry with pre-designed internal standard peptides specific to plant allergens, allowing for absolute and relative quantitation through spectral counting and peak integration, eliminating the need for gel electrophoresis and antibodies, and enabling the analysis of intact seeds without prior information.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If traditional methods (gel electrophoresis and antibodies) are used for allergen screening, then screening can be performed, but the process requires extensive resources and is inefficient
Solution Approach 1:
The patent replaces traditional mechanical and chemical methods (gel electrophoresis and antibody-based assays) with mass spectrometry technology. This substitution eliminates the need for extensive laboratory equipment, reagents, and manual operations, thereby improving screening efficiency while reducing resource requirements and complexity.
2Measurement precision
If traditional screening methods are used, then allergen detection can be performed, but accuracy in distinguishing natural and genetically-modified crops is insufficient
Solution Approach 1:
The patent employs stable isotope-labeled internal standard peptides with precisely known mass-to-charge ratios as reference points. By monitoring specific mass spectral peaks and comparing their intensities against these calibrated standards, the system achieves accurate absolute quantitation of allergen concentrations. This parameter-based approach enables reliable distinction between natural and genetically-modified crops based on precise allergen content measurement.
3Measurement precision
If comprehensive allergen screening is performed, then accurate quantitation is achieved, but analysis time and cost increase
Solution Approach 1:
The patent employs stable isotope-labeled internal standard peptides that are pre-synthesized and pre-characterized with known sequences, masses, and fragmentation patterns. These standards are prepared in advance and stored for reuse. During analysis, they are simply added to samples as internal references, eliminating the need for time-consuming sample preparation and calibration procedures, thereby achieving rapid absolute quantitation.
Solution Approach 2:
The mass spectrometry platform is designed to simultaneously analyze multiple allergen peptides in a single run. By using targeted multiple reaction monitoring (MRM) or parallel reaction monitoring (PRM) modes, the system can quantify numerous allergens across different plant sources concurrently, significantly reducing total analysis time while maintaining absolute quantitation accuracy for each target.
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 accurate, cost-effective, and rapid quantitation of plant seed allergens, capable of multiplexing multiple peptides in a single analysis, offering specific and absolute quantitation with reduced analysis time and cost.
Implementation Method 1
analyzing the sample using mass spectrometry with a set of pre-designed internal standard peptides that are specific for the plant allergen
Implementation Method 2
synthesized chemically with a heavy amino acid residue or genetically by expressing a synthetic gene in the presence of stable isotope-labeled amino acid(s) or metabolic intermediates
Data Source
AI summary
The invention provides a high-throughput method for quantitating plant seed proteins, e.g. seed allergens. Such method involves obtaining a protein sample from the seed; analyzing the sample using mass spectrometry with a set of pre-designed internal standard peptides that are specific to the plant allergens; and monitoring frequencies and intensities of resulting spectra to obtain relative and absolute allergen contents in the seed. The invention also provides a system for high-throughput profiling of plant seed allergens. Such system comprises a set of pre-designed internal standard peptides that are specific to the plant seed allergens and one or more mass spectrometers.