Isobaric Mass Labels for High-Plex Peptide Quantification
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Solution Overview
Problem
Current isobaric mass tags for multiplexing in mass spectrometry analysis are limited to 10-plex and face challenges with high complexity in mass spectra, making it difficult to achieve higher multiplexing rates without increasing tag size or complexity.
Innovation Solution
Development of a set of isotopomeric reactive mass labels with millidalton mass differences, allowing for higher multiplexing rates by using co-selectable isotopologue arrays that maintain the same integer mass but differ in exact mass, enabling more precise identification and quantification of peptides and biomolecules.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If traditional isobaric mass tags are used for multiplexing, then quantification capability is improved, but spectral complexity increases and multiplexing rate is limited to 10-plex
Solution Approach 1:
The mass tag is segmented into three functional components: a reporter ion (X) for identification, a cleavable bond (L) for release, and a mass normalizer (M) for isobaric matching. This segmentation allows each component to perform its specific function efficiently, enabling higher multiplexing without proportionally increasing overall complexity
Solution Approach 2:
The invention transitions from relying solely on integer mass differences to utilizing exact mass differences (millidalton level) as the differentiating dimension. This additional dimensional precision in mass measurement enables discrimination between more tags simultaneously, breaking the 10-plex limitation while maintaining spectral manageability
2Adaptability or versatility
If the number of mass tags is increased to achieve higher multiplexing, then quantification of more peptides is improved, but the complexity of mass spectra increases making identification difficult
Solution Approach 1:
The invention changes the critical parameter from integer mass difference to exact mass difference at the millidalton level. By utilizing high-resolution mass measurement capability, tags can be distinguished by extremely small mass variations, allowing many more tags to be resolved without increasing spectral complexity in a manageable way
3Measurement precision
If heavy isotope substitutions are used to create mass differences, then tag distinguishability is improved, but the mass difference becomes large (4-8 Daltons) limiting multiplexing rate
Solution Approach 1:
The invention changes the mass difference parameter from large (4-8 Daltons) to extremely small (millidalton level). This parameter change enables a much larger number of distinct tags to be created within a narrow mass window, directly increasing multiplexing rate while maintaining distinguishability through high-resolution mass measurement
Data Source
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AI summary
The present invention relates to a set of two or more mass labels, wherein each mass label comprises the formula: X-L-M-Re wherein X is a reporter moiety having an exact mass, L is a bond cleavable by collision in a mass spectrometer, M is a mass modifier, and Re is a) a reactive functionality for attaching the mass label to an analyte or b) the analyte, wherein each mass label in the set has an integer mass, wherein each mass label in the set has the same integer mass, and wherein the set comprises two or more subsets of mass labels, each subset comprising one, two or more mass labels, and wherein, when the subset comprises two or more mass labels, the exact mass of the reporter moiety X of each mass label in the subset is different from the exact mass of the reporter moiety X of the mass labels in the same subset and in all other subsets, and wherein each mass label is distinguishable by mass spectrometry.