Isotope-Coded Reporter Molecules for Multiplexed LC-MS/MS Detection
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Solution Overview
Problem
Conventional mass spectrometry (MS) assays are limited in their ability to simultaneously detect and quantify multiple analytes in complex biological samples due to variability in physiochemical properties, leading to challenges in sensitivity, specificity, and accuracy, especially in detecting multiple naturally occurring analytes with varying chemical structures and in complex samples like blood or tissue biopsies.
Innovation Solution
The development of isotope-coded reporter molecules (iCOREs) that are isobaric and distinguished by unique fragmentation ion signatures, allowing for the simultaneous detection of a virtually unlimited number of analytes through biochemical encoding and translation into mass-encoded reporters for MS-based detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If conventional MS assays are used to detect multiple analytes simultaneously, then the detection capability for multiple analytes is achieved, but the sensitivity, specificity, and accuracy are limited due to variability in physiochemical properties of different analytes
Solution Approach 1:
The patent introduces isotope-coded reporters (ICRs) as intermediary molecules that couple analytes to standardized mass tags. These ICRs serve as mediators between the diverse analytes and the MS detection system, translating varied analyte signals into uniform, highly detectable mass tag signals. This intermediary approach resolves the contradiction by decoupling the detection precision from analyte physiochemical variability.
Solution Approach 2:
The patent employs isotope labeling to change the mass-to-charge ratio parameter of analytes by attaching isotope-coded reporters. This parameter transformation converts analytes with varying physiochemical properties into standardized mass tags with distinct but predictable mass shifts, enabling precise simultaneous detection of multiple analytes without sensitivity compromise.
2Adaptability or versatility
If conventional MS assays monitor a wide mass window to detect multiple analytes with varying chemical structures, then the detection coverage is improved, but the analysis time increases significantly
Solution Approach 1:
The patent transforms analyte masses into a standardized mass window by attaching isotope-coded reporters with known mass shifts. Instead of monitoring a wide mass window covering all possible analyte masses, the system monitors a narrow, predictable mass window where all analyte-ICR conjugates appear, dramatically reducing analysis time while maintaining comprehensive detection coverage.
Solution Approach 2:
The isotope-coded reporters are designed with predetermined mass shifts, allowing the system to anticipate and target specific mass regions for detection. This preliminary structuring of mass distributions enables focused monitoring of narrow mass windows, eliminating the need for time-consuming broad-spectrum scans.
3Adaptability or versatility
If conventional MS assays are used for detecting target analytes in complex biological samples, then the detection of endogenous analytes is achieved, but extensive prior front-end processing is required
Solution Approach 1:
The isotope-coded reporters act as intermediaries that simplify the detection of endogenous analytes in complex biological samples. By coupling analytes to standardized ICRs with distinct mass signatures, the system enhances analyte detectability and enables straightforward differentiation from background noise, reducing the need for complex pre-processing steps.
Solution Approach 2:
The attachment of isotope-coded reporters changes the mass parameter of endogenous analytes, creating detectable mass shifts that distinguish target analytes from the complex biological matrix. This parameter transformation simplifies sample processing by enabling direct detection without extensive purification or enrichment steps.
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 sensitive, specific, and accurate multiplexed detection and quantification of analytes, reducing the need for extensive pre-processing and increasing detection sensitivity by 30-300 fold, while simplifying data collection and reducing analysis time and costs, far exceeding the multiplexing capabilities of current MS technologies.
Implementation Method 1
detectable molecules, called mass tags, are ionized to generate charged molecules or molecule fragments and subsequently the mass-to-charge ratio of these molecules is measured
Implementation Method 2
different fragmentation signatures are conferred to different iCOREs by differential isotope labeling
Data Source
AI summary
Some aspects of this invention provide reagents and methods for the sensitive, quantitative and simultaneous detection of target analytes in complex biological samples by liquid chromatography tandem mass spectrometry (LC MS/MS). Some aspects of this invention provide affinity reagents encoded with mass reporters for the sensitive and quantitative translation of an analyte of interest into a mass tag. The reagents and methods provided herein have general utility in analyte detection and encoding, for example, in biomolecular profiling, molecular diagnostics, and biochemical encoding.