Concurrent analysis of multiple analytes
AssigneeThe Regents of the University of California
InventorsAustin QUACH, Kym Francis FAULL
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Abstract
Methods, apparatuses and systems are described that are capable of simultaneously determining the presence, identities or levels of multiple analytes present in a single sample, by carrying out steps including denaturation, normalization, extraction, mixed-mode liquid chromatography and mass spectrometry, whereby the presence, identities or levels of analytes in the single sample are determined.
Claims100
A method for substantially simultaneously determining the presence, identities or amounts of multiple analytes present in a single sample, the method comprising:
- a. treating the sample with a denaturation reagent or treatment that denatures biopolymers in the sample thereby forming a denatured sample, wherein the denaturation reagent or treatment does not denature components of a normalization reagent when added thereto;
- b. treating the denatured sample with the normalization reagent, wherein the normalization agent converts the multiple analytes in the denatured sample into normalized analyte species and normalized analyte fragment species, the species capable of being separated by multiple modes of chromatography and individually identified by mass spectrometry, thereby forming a normalized sample;
- c. treating the normalized sample with an extraction reagent and retaining the soluble analyte species and analyte fragment species therein, thereby forming an extracted sample;
- d. subjecting the extracted sample to multiple modes of chromatography followed by mass spectrometry (MS), whereby data therefrom for each normalized analyte species and normalized analyte fragment species present therein are generated;
- and
- e. computationally determining from the data the presence, identities or amounts of the multiple analytes present in the extracted sample.
An apparatus for substantially simultaneously determining the presence, identities or amounts of multiple analytes present in a single sample, the apparatus comprising:
- a. means for denaturing biopolymers in the sample thereby forming a denatured sample, wherein the denaturing does not denature components of a normalization reagent when added thereto;
- b. means for normalizing the sample wherein multiple analytes in the denatured sample are converted into normalized analyte species and normalized analyte fragment species, the species capable of being separated by multiple modes of chromatography and individually identified by mass spectrometry;
- c. means for extracting the normalized sample and retaining the soluble analyte species and analyte fragment species therein;
- d. means for subjecting the extracted sample to multiple modes of chromatography followed by mass spectrometry, whereby data therefrom for each normalized analyte species and normalized analyte fragment species present therein are generated;
- and
- e. means for computationally determining from the data the presence, identities or amounts of the multiple analytes present in the extracted sample.
A system for substantially simultaneously determining the presence, identities or amounts of multiple, chemically-related and chemically-unrelated analytes present in a single sample, the system comprising:
- a. a denaturation reagent or treatment that denatures biopolymers in the sample thereby forming a denatured sample, wherein the denaturation reagent or treatment does not denature components of a normalization reagent when added thereto;
- b. a normalization reagent that converts the multiple analytes in the sample into normalized analyte species or normalized analyte fragment species capable of being separated by mixed mode liquid chromatography and individually identified by tandem mass spectrometry;
- c. an extraction reagent that extracts the soluble analyte species and analyte fragment species;
- d. a separation process for retaining the soluble analyte species and analyte fragment species therein, thereby forming an extracted sample;
- e. multiple modes of chromatography that resolves soluble analyte species and analyte fragment species;
- f. mass spectrometry that generates data on the individual species;
- and
- g. one or more algorithms for computationally determining from the data of each normalized analyte species and normalized analyte fragment species the presence, identities or amounts of the multiple analytes present in the sample.
A method for identifying a plurality of analytes of different types from a sample, comprising:
- (a) subjecting said sample containing or suspected of containing said plurality of analytes of different types to conditions sufficient to yield a solution comprising said plurality of analytes or derivatives thereof;
- and
- (b) using an instrument to process said solution to identify said plurality of analytes or derivatives thereof, thereby identifying said plurality of analytes, wherein said plurality of analytes or derivatives thereof is identified in a single run of said instrument.
A method for determining a disease or condition in a subject, comprising:
- (i) identifying a plurality of analytes, individually or collectively, known to be associated with said disease or condition from a single sample of said subject to obtain an identified amount for each of said plurality of analytes, wherein said plurality of analytes comprises analytes of different types;
- (ii) determining a difference between a reference amount and said identified amount for said each of said plurality of analytes to obtain a plurality of difference values;
- and
- (iii) using a trained machine learning algorithm to determine said disease or condition based on said plurality of difference values.
Dependencies are inferred from the claim text.