Isobaric Labeling Reagents for Multiplex Mass Spectrometry
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Solution Overview
Problem
Current methods for analyte determination by mass analysis face challenges in distinguishing between isobaric samples and accurately quantifying analytes in complex mixtures, particularly in multiplex analyses where samples are mixed and analyzed together.
Innovation Solution
The use of isomeric or isobaric labeling reagents with unique reporter moieties that fragment differently upon dissociative energy, allowing for the identification and quantification of analytes based on their distinct mass signatures in mass spectrometry, even when samples are mixed and analyzed together.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If isobaric labeling reagents are used to label analytes from different samples, then multiplex analysis capability is improved, but the ability to distinguish between isobaric samples deteriorates
Solution Approach 1:
The labeling reagent is divided into two functional segments: a common linker portion and sample-specific reporter portions. The reporter portions are designed to be chemically identical but have different masses, allowing them to be segmented by mass in the mass spectrometer while maintaining chemical uniformity for multiplex analysis.
Solution Approach 2:
The invention changes the mass parameter of the reporter portions while keeping chemical properties identical. By using isotopic labeling (e.g., 13C, 15N, 18O) to create mass differences of 1-10 Daltons between reporter portions, the system enables distinction between samples through mass analysis while maintaining chemical equivalence for labeling purposes.
2Measurement precision
If unique reporter moieties with different masses are used for each sample, then sample identification accuracy is improved, but the complexity of the labeling reagent system increases
Solution Approach 1:
The linker portion serves multiple functions: it provides identical chemical reactivity for labeling analytes from different samples, acts as a common structural framework, and enables multiplex analysis capability. This universal component reduces system complexity while the variable reporter portions provide sample-specific identification.
Solution Approach 2:
The reporter portions are designed with local quality differences (different masses) while maintaining the same overall chemical structure and reactivity. This allows sample-specific identification through mass differences without requiring fundamentally different chemical chemistries, thereby limiting the increase in system complexity.
3Measurement precision
If isobaric labeling reagents are used, then quantification accuracy in complex mixtures is improved, but the difficulty of detecting and measuring analytes deteriorates
Solution Approach 1:
The reporter portions act as intermediary structures that transfer the mass information to the analyte during labeling. After the analyte is labeled, the reporter portions can be selectively removed or fragmented, allowing the mass spectrometer to detect the analyte's mass shifted by the reporter's mass, thereby facilitating accurate quantification without direct interference with the analyte's detection.
Solution Approach 2:
The invention adds a mass dimension to the detection process by using reporter portions with controlled mass differences. This allows the mass spectrometer to distinguish between analytes from different samples based on their total mass (analyte + reporter), providing an additional dimension of information for accurate quantification in complex mixtures.
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
Enables precise identification and quantification of analytes in complex mixtures by leveraging the unique fragmentation patterns of isomeric or isobaric labeling reagents, facilitating multiplex analysis and improving the accuracy of analyte determination in mass spectrometry.
Implementation Method 1
the labeling reagent can comprise a unique reporter, 'RP', that can be associated with the sample from which the labeled analyte originated... the reporter can be identified with, and therefore used to identify, the sample from which each labeled analyte originated
Data Source
AI summary
This invention pertains to methods, mixtures, kits and/or compositions for the determination of analytes by mass analysis using unique labeling reagents or sets of unique labeling reagents. The labeling reagents can be isomeric or isobaric and can be used to produce mixtures suitable for multiplex analysis of the labeled analytes.