Isobaric mass labels
Inventors
Thompson, Andrew Hugin • Kienle, Stefan • Kuhn, Karsten • Woellmer, Nikolai
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-10527629-B2
Publication Date
2020-01-07
Expiration Date
Abstract
The present invention relates to a set of two or more mass labels. Each mass label comprises: X-L-M-Re; X is a reporter moiety having exact mass; L is a bond cleavable by collision in a mass spectrometer; M is a mass modifier; Re is a) a reactive functionality for attaching the mass label to an analyte or b) the analyte. Each mass label in the set has the same integer mass; wherein the set comprises two or more subsets of mass labels, each subset comprising one or more mass labels; 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; each mass label is distinguishable by mass spectrometry.
Core Innovation
The invention relates to sets of two or more mass labels for use in a mass spectrometer, in which each label comprises X-L-M-Re, where 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 reactive functionality for attaching the mass label to an analyte or the analyte itself. The reporter moiety X is defined by substituted or unsubstituted structures with specified substituents, including straight or branched C1-C10 alkyl, aliphatic cyclic, aromatic, heterocyclic, and hydrogen options.
The disclosure uses isobaric, isotopologue, and pseudo-isobaric mass label set designs to support multiplexing and correct reporter-ion assignment. Heavy isotope substitution, including 13C, 15N, 18O, and 2H, marks substitutable positions, and the labels are organized into sets, subsets, and arrays with distinguishable reporter exact masses and integer-mass relationships.
The document also describes mass spectrometry analysis workflows in which analyte-containing samples are differentially labeled, mixed, dissociated to generate intact mass labels and/or analyte fragments, and identified from the resulting mass spectra. The label sets support co-elution, co-selection, minimal mobility shift, and reporter-ion assignment, with high-resolution MS instruments such as Orbitrap, FTICR, TOF, and MALDI TOF mentioned in the disclosed analysis context.
Claims Coverage
The consolidated claim coverage centers on the recurring independent concept of a set of two or more mass labels defined by the X-L-M-Re structure and a constrained reporter moiety X. Across the provided items, six inventive features are represented, including label structure, reporter moiety definition, isotope-restricted compositions, subset distinguishability, array-level integer-mass non-overlap, and a differential-labeling detection workflow.
Mass label set with X-L-M-Re structure
A set of two or more mass labels, each label comprising X-L-M-Re, where 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 reactive functionality for attaching the mass label to an analyte or the analyte itself.
Reporter moiety structure for mass label set X
X comprises substituted or unsubstituted straight or branched C1-C10 alkyl, aliphatic cyclic, aromatic, or heterocyclic structures, with defined selections for R1, R2, R3, R4, R5, R6, and R9.
Integer-mass distinguishability using subsets
A set of mass labels in which each label is distinguished by mass spectrometry with an integer mass, and the set includes two or more subsets having different reporter moiety X exact masses, with different exact masses between labels within a subset and across subsets.
Isotope-restricted mass label compositions
A set of mass labels in which labeled atoms are restricted to 13C or 15N, including examples of a five-label set and an eighteen-label set using only 13C or 15N.
Array design with integer mass non-overlap
An array of mass labels in which, within the array, the integer mass values in any one set are all different from the integer mass values in every other set.
Differential labeling and dissociation workflow for analyte identification
A method that detects one or more analytes by mass spectrometry by differentially labeling analyte-containing samples with defined mass labels, mixing labeled samples, dissociating to generate intact mass labels and/or analyte fragments, detecting those intact labels/fragments, and identifying analytes from the resulting mass spectra.
The claim coverage centers on the X-L-M-Re mass-label architecture with a defined reporter moiety X, together with isotope, subset, and array constraints that support mass-spectrometric distinguishability. The covered workflow additionally uses differential labeling and dissociation to identify analytes from mass spectra.
Stated Advantages
Enables multiplexing through isobaric, isotopologue, and pseudo-isobaric mass label set designs.
Supports reporter-ion assignment in multiplexed contexts despite co-elution and co-selection considerations.
Provides correct reporter-ion assignment through co-selectable parent tag masses with different reporter moiety masses.
Allows co-elution, co-selection, and minimal mobility shift.
Supports high multiplexing capacities.
Documented Applications
Multiplexing using isobaric, isotopologue, and pseudo-isobaric mass label sets for mass spectrometry analysis.
Differential labeling, mixing, dissociation, and detection or identification from mass spectra.
Design and generation of arrays and subsets of mass labels using integer-mass non-overlap and isotopologue relationships.
High-resolution MS-based analysis using Orbitrap, FTICR, TOF, and MALDI TOF for detecting intact mass labels and reporter moieties.
Interested in licensing this patent?