Isobaric Label Fragmentation Energy for Mass Spectrometry
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current isobaric labels in mass spectrometry analysis cannot selectively fragment at lower energies without fragmenting analytes, limiting the throughput and spectral resolution, especially when coupled with devices like liquid chromatography, as they require high energies to fragment both the label and the peptide analyte, leading to inefficient use of time on the mass spectrometer.
Innovation Solution
Development of an isobaric label that fragments at energies below those required for peptide fragmentation, with a higher conversion rate, allowing for selective fragmentation of the label while keeping the peptide intact, thereby enabling more efficient data-independent acquisition and improved spectral resolution.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If high energy is used to fragment isobaric labels, then label fragmentation is achieved, but analyte (peptide) fragmentation also occurs and time on the mass spectrometer is wasted
Solution Approach 1:
The isobaric label is designed with a specific chemical structure containing a labile bond that fragments at lower collision energies than required for peptide backbone fragmentation. This parameter change in fragmentation energy threshold allows selective label fragmentation without analyte degradation, improving spectral resolution while reducing time waste on the mass spectrometer.
2Reliability
If high energy is used to fragment both label and peptide, then complete fragmentation is achieved, but throughput decreases due to inefficient use of mass spectrometer time
Solution Approach 1:
The fragmentation process is extracted and isolated to only the isobaric label portion of the conjugate, leaving the peptide analyte intact. This is achieved through the specific chemical design of the label containing a low-energy lable bond that breaks preferentially, enabling complete label fragmentation while preserving the analyte for further analysis, thus improving throughput without sacrificing reliability.
3Strength
If high energy fragmentation is used, then both label and analyte are fragmented, but spectral resolution and throughput are limited
Solution Approach 1:
The isobaric label is designed with localized chemical features (labile bonds, specific functional groups) that create a spatial and energetic distinction between label and analyte fragmentation. The label portion has optimized local chemical properties enabling low-energy fragmentation, while the peptide analyte maintains its structural integrity, achieving high fragmentation efficiency for the label without compromising overall productivity.
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 allows for better utilization of time on the mass spectrometer by enabling the use of lower energies for label fragmentation, increasing throughput and spectral resolution, and enabling the analysis of multiple precursor ions in a single MS2 scan, thereby improving the identification and quantification of peptides in complex mixtures.
Implementation Method 1
said isobaric label comprises or consists of a group which fragments in the mass spectrometer (i) at an energy below the energy required for fragmenting analyte-derived precursor ions and/or a higher conversion rate than said precursor ions
Data Source
AI summary
The present invention relates to use of an isobaric label in mass spectrometry (MS) analysis using data-independent acquisition (DIA), wherein said isobaric label comprises or consists of a group which fragments in the mass spectrometer (i) at an energy below the energy required for fragmenting analyte-derived precursor ions and/or a higher conversion rate than said precursor ions; and (ii) at said energy according to (i) and when coupled to a precursor ion, at a single site within said group, to yield a first moiety and a second moiety, said second moiety being coupled to said precursor ion.