Back to results
US 2011/0297823 A1Published application
Mass spectrometry data acquisition mode for obtaining more reliable protein quantitation
AssigneeWisconsin Alumni Research Foundation
InventorsJoshua J. Coon, Douglas H. Phanstiel, Craig D. Wenger
Life of this patent
Estimating the term…
Abstract
Described herein are methods and systems which enable a unique platform for analyte quantitation. The methods and systems relate to determining the amount of interference in a precursor ion isolation window resulting from an impurity. Once the level of impurity is determined, several methods can be employed to reduce the amount of interference in a subsequent MS/MS spectrum. The methods and systems described herein enable increased quantitation accuracy while maintaining high levels of throughput.
Claims26
A method of analyzing an analyte using mass spectrometry, the method comprising:
- (a) providing an analyte;
- (b) generating a distribution of precursor ions from the analyte;
- (c) analyzing the mass-to-charge ratios of at least a portion of the distribution of precursor ions, thereby generating precursor ion mass spectrometry data corresponding to the distribution of precursor ions;
- (d) identifying a precursor peak in the precursor ion mass spectrometry data corresponding to a precursor ion;
- (e) fragmenting ions corresponding to a preselected range of m/z units about the precursor peak, wherein the preselected range is within 0.01 to 10 m/z units of the precursor peak, thereby generating fragment ions;
- (f) measuring the mass-to-charge ratios of the fragment ions, thereby generating product ion mass spectrometry data;
- (g) determining the amount of interference within the preselected range of m/z units about the precursor peak;
- and
- (h) analyzing the precursor ion mass spectrometry data and the product ion mass spectrometry data for which the amount of interference is less than a selected value, and not analyzing the precursor ion mass spectrometry data and the product ion mass spectrometry data for which the amount of interference is greater than or equal to the selected value;
- thereby analyzing the analyte using mass spectrometry.
7 more elements
A method of analyzing an analyte using mass spectrometry, the method comprising:
- (a) providing an analyte;
- (b) generating a distribution of precursor ions from the analyte;
- (c) analyzing the mass-to-charge ratios of at least a portion of the distribution of precursor ions, thereby generating precursor ion mass spectrometry data corresponding to the distribution of precursor ions;
- (d) identifying a precursor peak in the precursor ion mass spectrometry data corresponding to a precursor ion;
- (e) determining the amount of interference within a preselected range of m/z units about the precursor peak, wherein the preselected range is within 0.01 to 10 m/z units of the precursor peak;
- (f) fragmenting ions corresponding to the preselected range of m/z units about the precursor peak when the amount of interference is less than a selected value, thereby generating fragment ions;
- and not fragmenting ions corresponding to the preselected range of m/z units about the precursor peak when the amount of interference is greater than or equal to the selected value;
- (g) measuring the mass-to-charge ratios of the fragment ions, thereby generating product ion mass spectrometry data;
- and
- (h) analyzing the precursor ion mass spectrometry data and the product ion mass spectrometry data;
- thereby analyzing the analyte using mass spectrometry.
8 more elements
A method of analyzing an analyte using mass spectrometry, the method comprising:
- (a) providing an analyte;
- (b) generating a distribution of precursor ions from the analyte;
- (c) analyzing the mass-to-charge ratios of at least a portion of the distribution of precursor ions, thereby generating precursor ion mass spectrometry data corresponding to the distribution of precursor ions;
- (d) identifying a precursor peak in the precursor ion mass spectrometry data corresponding to a precursor ion;
- (e) determining the amount of interference within a preselected range of m/z units about the precursor peak, wherein the preselected range is within 0.01 to 10 m/z units of the precursor peak;
- (f) fragmenting ions corresponding to the preselected range, thereby generating fragment ions;
- (g) measuring mass-to-charge ratios of fragment ions corresponding to a preselected range when the amount of interference is less than a selected value, and not measuring mass-to-charge ratios of fragment ions corresponding to a preselected range when the amount of interference is greater than or equal to the selected value, thereby generating product ion mass spectrometry data;
- and
- (h) analyzing the precursor ion mass spectrometry data and the product ion mass spectrometry data;
- thereby analyzing the analyte using mass spectrometry.
7 more elements
A mass spectrometer system for analyzing an analyte, the system comprising:
- an ion source for generating ions from the analyte;
- first ion separation optics in communication with the ion source for separating ions according to their mass-to-charge ratios;
- a first ion detector in communication with the first ion separation optics for detecting ions separated according to their mass-to-charge ratios;
- ion fragmentation optics in communication with the first ion separation optics for generating fragment ions;
- second ion separation optics in communication with the ion fragmentation optics for separating ions according to their mass-to-charge ratios;
- a second ion detector in communication with the second ion separation optics for detecting ions separated according to their mass-to-charge ratios;
- a controller operably connected to the first and second ion separation optics, the first and second ion detectors, and the ion fragmentation optics;
- wherein the controller controls the ion optics and detectors so as to:
- (a) generate a distribution of precursor ions from the analyte;
- (b) analyze the mass-to-charge ratios of at least a portion of the distribution of precursor ions, thereby generating precursor ion mass spectrometry data corresponding to the distribution of precursor ions;
- (c) identify a precursor peak in the precursor ion mass spectrometry data corresponding to a precursor ion;
- (d) determine the amount of interference within a preselected range of m/z units about the precursor peak, wherein the preselected range is within 0.01 to 10 m/z units of the precursor peak;
- (e) fragment the ions corresponding to the preselected range, thereby generating fragment ions;
- (f) measure mass-to-charge ratios of fragment ions corresponding to a preselected range when the amount of interference is less than a selected value, and not measure mass-to-charge ratios of fragment ions corresponding to a preselected range when the amount of interference is greater than or equal to the selected value, thereby generating product ion mass spectrometry data;
- and
- (g) analyze the precursor ion mass spectrometry data and the product ion mass spectrometry data.
13 more elements
A mass spectrometer system for analyzing an analyte, the system comprising:
- an ion source for generating ions from the analyte;
- first ion separation optics in communication with the ion source for separating ions according to their mass-to-charge ratios;
- a first ion detector in communication with the first ion separation optics for detecting ions separated according to their mass-to-charge ratios;
- ion fragmentation optics in communication with the first ion separation optics for generating fragment ions;
- second ion separation optics in communication with the ion fragmentation optics for separating ions according to their mass-to-charge ratios;
- a second ion detector in communication with the second ion separation optics for detecting ions separated according to their mass-to-charge ratios;
- a controller operably connected to the first and second ion separation optics, the first and second ion detectors, and the ion fragmentation optics;
- wherein the controller controls the ion optics and detectors so as to:
- (a) generate a distribution of precursor ions from the analyte;
- (b) analyze the mass-to-charge ratios of at least a portion of the distribution of precursor ions, thereby generating precursor ion mass spectrometry data corresponding to the distribution of precursor ions;
- (c) identify a precursor peak in the precursor ion mass spectrometry data corresponding to a precursor ion;
- (d) fragment ions corresponding to a preselected range of m/z units about the precursor peak, wherein the preselected range is within 0.01 to 10 m/z units of the precursor peak, thereby generating fragment ions;
- (e) measure the mass-to-charge ratios of the fragment ions, thereby generating product ion mass spectrometry data;
- (f) determine the amount of interference within the preselected range of m/z units about the precursor peak;
- and
- (g) analyze the precursor ion mass spectrometry data and the product ion mass spectrometry data for which the amount of interference is less than a selected value, and not analyze the precursor ion mass spectrometry data and the product ion mass spectrometry data for which the amount of interference is greater than or equal to the selected value.
13 more elements
A mass spectrometer system for analyzing an analyte, the system comprising:
- an ion source for generating ions from the analyte;
- first ion separation optics in communication with the ion source for separating ions according to their mass-to-charge ratios;
- a first ion detector in communication with the first ion separation optics for detecting ions separated according to their mass-to-charge ratios;
- ion fragmentation optics in communication with the first ion separation optics for generating fragment ions;
- second ion separation optics in communication with the ion fragmentation optics for separating ions according to their mass-to-charge ratios;
- a second ion detector in communication with the second ion separation optics for detecting ions separated according to their mass-to-charge ratios;
- a controller operably connected to the first and second ion separation optics, the first and second ion detectors, and the ion fragmentation optics;
- wherein the controller controls the ion optics and detectors so as to:
- (a) generate a distribution of precursor ions from the analyte;
- (b) analyze the mass-to-charge ratios of at least a portion of the distribution of precursor ions, thereby generating precursor ion mass spectrometry data corresponding to the distribution of precursor ions;
- (c) identify a precursor peak in the precursor ion mass spectrometry data corresponding to a precursor ion;
- (d) determine the amount of interference within a preselected range of m/z units about the precursor peak, wherein the preselected range is within 0.01 to 10 m/z units of the precursor peak;
- (e) fragment ions corresponding to the preselected range when the amount of interference is less than a selected value, thereby generating fragment ions;
- and not fragment ions corresponding to the preselected range when the amount of interference is greater than or equal to the selected value;
- (f) measure mass-to-charge ratios of the fragment ions, thereby generating product ion mass spectrometry data;
- and
- (g) analyze the precursor ion mass spectrometry data and the product ion mass spectrometry data.
14 more elements
A method of analyzing an analyte using mass spectrometry, the method comprising:
- (a) providing an analyte;
- (b) generating a distribution of precursor ions from the analyte;
- (c) analyzing the mass-to-charge ratios of at least a portion of the distribution of precursor ions, thereby generating precursor ion mass spectrometry data corresponding to the distribution of precursor ions;
- (d) identifying a precursor peak in the precursor ion mass spectrometry data corresponding to a precursor ion;
- (e) determining an amount of interference within a range of 0.01 to 10 m/z units of the precursor peak;
- (f) adjusting the range of m/z units such that the amount of interference is less than a selected value;
- and
- (g) analyzing the ions within the adjusted range of m/z units;
- thereby analyzing the analyte using mass spectrometry.
6 more elements
Dependencies are inferred from the claim text.