Methods for conducting metabolic analyses
AssigneeTarget Discovery, Inc.
InventorLuke V. Schneider
Life of this patent
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Abstract
The present invention provides methods and apparatus for purifying metabolites of interest and conducting metabolic analyses. The methods generally involve determining metabolic flux values for a plurality of target analytes by monitoring the relative isotope abundance of a stable isotope in a substrate labeled with the stable isotope and/or one or more target metabolites formed through metabolism of the labeled substrate. Certain methods utilize multiple electrophoretic methods to separate the target analytes from other components within the sample being analyzed. The methods can be used in a variety of applications including screens to identify metabolites that are correlated with certain diseases and diagnostic screens for identifying individuals having, or susceptible to, a disease.
Claims35
A method for analyzing metabolic pathways, comprising:
- (a) administering to a subject a substrate labeled with a stable isotope, wherein the relative isotopic abundance of the isotope in the substrate is known;
- (b) allowing the labeled substrate to be at least partially metabolized by the subject to form one or more target metabolites;
- and
- (c) determining the abundance of the isotope in a plurality of target analytes in a sample from the subject to determine a value for the flux of each target analyte, wherein the plurality of target analytes comprise the substrate and/or one or more of the target metabolites.
A method for analyzing metabolic pathways, comprising:
- (a) separating at least partially a plurality of target analytes from biological components contained in a sample obtained from a subject, the target analytes comprising a substrate labeled with a stable isotope and/or one or more target metabolites resulting from the metabolism of the substrate by the subject, and wherein the relative isotopic abundance of the isotope in the substrate is known;
- and
- (b) determining the abundance of the isotope in a plurality of the target analytes in the sample to determine a value for the flux of each target analyte.
A method for screening for metabolites correlated with a disease, comprising:
- (a) administering to a test subject and a control subject a substrate labeled with a stable isotope, wherein the relative isotopic abundance of the isotope in the substrate is known and the test subject has the disease;
- (b) allowing the labeled substrate to be at least partially metabolized by the test subject and control subject to form one or more target metabolites, and wherein the conditions under which the administering and allowing steps are performed the same for the test and control subject;
- and
- (c) obtaining a sample from the test and control subject;
- (d) determining for each sample the relative abundance of the isotope in a plurality of target analytes to determine a value for the flux of each target analyte, wherein the target analytes comprise the substrate and/or one or more of the target metabolites;
- and
- (e) comparing the values for flux for the test and control subjects, a difference in the flux value for a target analyte in the test subject and corresponding flux value for the control subject indicating that such analyte is potentially correlated with the disease.
A method for screening for metabolites correlated with a disease, comprising:
- (a) analyzing a sample from a test subject having the disease, the sample comprising a substrate labeled with a stable isotope administered to the test subject and/or one or more target metabolites resulting from metabolism of the substrate by the test subject, the relative isotopic abundance of the isotope in the substrate known at the time of administration, and wherein the analyzing step comprises determining the isotopic abundance of the isotope in a plurality of analytes in the sample to determine a value for the flux of each analyte, wherein the plurality of analytes comprise the substrate and/or one or more of the target metabolites;
- and
- (b) comparing flux values for the analytes with flux values for the same analytes obtained for a control subject, wherein a difference in a flux value for an analyte indicates that such analyte is correlated with the disease.
A method for screening for the presence of a disease, comprising:
- (a) administering to a test subject a substrate labeled with a stable isotope, wherein the relative abundance of the isotope in the substrate is known;
- (b) allowing sufficient time for the labeled substrate to be at least partially metabolized by the test subject to form one or more target metabolites known to be correlated with the disease;
- (c) performing a plurality of electrophoretic methods in series to at least partially separate a plurality of target analytes from other biological components in a sample obtained from the test subject, wherein the target analytes comprise the substrate and/or one or more of the target metabolites;
- (d) determining a flux value for the target analytes, the flux value for each target analyte being determined from the abundance of the isotope in that analyte;
- and
- (e) comparing determined flux values with corresponding reference flux values for the same target analytes to assess the test subject's risk of disease.
A method for screening for the presence of a disease, comprising:
- (a) analyzing a sample from a test subject, the sample comprising a substrate labeled with a stable isotope administered to the test subject and/or one or more target metabolites resulting from metabolism of the substrate by the test subject, the relative isotopic abundance of the isotope in the substrate known at the time of administration, and wherein the analyzing step comprises determining the abundance of the isotope in a plurality of analytes in the sample to determine a value for the flux of each analyte, wherein the plurality of analytes comprise the substrate and/or one or more of the target metabolites;
- and
- (b) for each target analyte, comparing the determined flux value with a range of flux values for that analyte, wherein the range is known to be correlated with the disease and a determined flux value for a target analyte falling within the range for that target analyte, indicates that the test subject has the disease or is susceptible to the disease.
Dependencies are inferred from the claim text.