Mass spectrometric analysis of complex mixtures of immune system modulators
AssigneeThe Johns Hopkins University
InventorJoany Jackman
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Abstract
The invention is directed to methods suitable for detecting the presence of a plurality of immune system modulators in a complex solution by running a single sample through a mass spectrometer without any surface preparation or preliminary purification step. Immune system modulators that can be analyzed with the methods of the present invention include cytokines (interferons and interleukins), chemokines, host cell response molecules, and peptide fragments. The method also allows the identification of truncated receptors for immune modulators secreted by infected cells or pathogens, such as virokines, and other pathogen-encoded proteins that affect the host immune system.
Claims24
A method for analyzing the presence of one or more immune system modulators in a complex mixture using mass spectrometry, comprising the steps of:
- a. obtaining a complex mixture for mass spectral analysis, wherein the complex mixture includes exhaled breath condensate,
- b. placing a sample of the complex mixture including the exhaled breath condensate on a substratum suitable for use in a mass spectrometer,
- c. saturating the sample with matrix material,
- d. allowing the sample to dry,
- e. inserting the sample into a mass spectrometer, and
- f. processing the sample in the mass spectrometer to obtain a mass spectrograph of the complex mixture including the exhaled breath condensate, wherein said processing includes determining, from the mass spectrograph, whether the sample of exhaled breath condensate includes one or more immune system modulators comprising at least a fragment of a protein.
A method for analyzing the presence of one or more molecular markers of infection in a complex mixture using mass spectrometry, comprising the steps of:
- a. obtaining a complex mixture for mass spectral analysis, wherein the complex mixture includes exhaled breath condensate,
- b. placing a sample of the complex mixture including the exhaled breath condensate on a substratum suitable for use in a mass spectrometer,
- c. saturating the sample with matrix material,
- d. allowing the sample to dry,
- e. inserting the sample into a mass spectrometer, and
- f. processing the sample in the mass spectrometer to obtain a mass spectrograph of the complex mixture including the exhaled breath condensate,
- wherein the method further includes
- obtaining a known sample of pure molecular marker of infection standard for a particular molecular marker of infection comprising at least a fragment of a protein,
- placing the known sample on a substratum suitable for use in a mass spectrometer,
- repeating steps c–f for the known sample to obtain a mass spectrograph of the pure molecular marker of infection standard,
- comparing the mass spectrograph of the complex mixture to the mass spectrograph of the pure molecular marker of infection standard, and
- determining whether the complex mixture includes the particular molecular marker of infection, comprising at least a fragment of a protein, based on whether one or more peaks of the mass spectrograph of the pure molecular marker of infection standard are identified in the mass spectrograph of the complex mixture.
A method for analyzing the presence of one or more immune system modulators in a complex mixture using mass spectrometry, comprising the steps of:
- a. obtaining a complex mixture for mass spectral analysis, wherein the complex mixture includes exhaled breath condensate having the one or more immune system modulators therein,
- b. placing a sample of the complex mixture including the exhaled breath condensate on a substratum suitable for use in a mass spectrometer,
- c. saturating the sample with matrix material,
- d. allowing the sample to dry,
- e. inserting the sample into a mass spectrometer, and
- f. processing the sample in the mass spectrometer to obtain a mass spectrograph of the complex mixture including the exhaled breath condensate, wherein the one or more immune system modulators includes multiple immune system modulators, and wherein step f. comprises:
- obtaining multiple spectrographs each corresponding to a different one of the multiple system modulators;
- normalizing each of the multiple spectrographs to a common maximum ionization signal among the multiple spectrographs;
- and
- overlaying and aligning the normalized multiple spectrographs.
Dependencies are inferred from the claim text.