Radioactive multiplexing analytical methods for biomarkers discovery
First Claim
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1. A method of analysis comprising the steps of:
- (a) labeling molecules in a first sample with a first radioactive isotope;
(b) labeling molecules in a second sample with a second radioactive isotope that is different from said first radioactive isotope;
(c) mixing said first sample and said second sample into a mixture;
(d) separating said mixture into a plurality of groups of molecules; and
,(e) using the difference in half-life between said first radioactive isotope and said second radioactive isotope as a means to selectively quantify each isotope in each group of molecules.
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Abstract
A novel analytical method involves labeling samples with different radioactive labeling agents, mixing and subjecting the mixture to any separation technique, and then differentially detecting and quantifying subcomponents from each sample for comparison. The novel technique exploits the differences in radiation energy or half-life between isotopes to make differential detection and quantification of labels possible. Detailed methods for differential detection and quantification are also described as well as the construction and application of hardware and software to enable and enhance such a process. This method is useful in finding molecular differences between two samples in differential proteomics, phosphor-proteomics, glycomics, metabolomics, transcriptomics, genomics and diagnostic applications.
12 Citations
17 Claims
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1. A method of analysis comprising the steps of:
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(a) labeling molecules in a first sample with a first radioactive isotope; (b) labeling molecules in a second sample with a second radioactive isotope that is different from said first radioactive isotope; (c) mixing said first sample and said second sample into a mixture; (d) separating said mixture into a plurality of groups of molecules; and
,(e) using the difference in half-life between said first radioactive isotope and said second radioactive isotope as a means to selectively quantify each isotope in each group of molecules. - View Dependent Claims (3, 4, 5)
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2. A method of analysis comprising the steps of:
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(a) labeling molecules in a first sample with a first radioactive isotope; (b) labeling molecules in a second sample with a second radioactive isotope that is different from said first radioactive isotope; (c) mixing said first sample and said second sample into a mixture; (d) separating said mixture into a plurality of groups of molecules; and
,(e) selectively quantifying each isotope from each group of molecules wherein a screen is used to selectively block a higher proportion of radiation from one isotope.
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6. A method of isolating a difference between two samples comprising the steps of:
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(a) labeling molecules in a first sample with a first radioactive isotope; (b) labeling molecules in a second sample with a second radioactive isotope that is different from said first radioactive isotope; (c) mixing said first sample and said second sample into a mixture; (d) separating said mixture into a plurality of fractions; (e) quantifying a ratio of radioactive isotopes for each fraction; and
,(f) comparing the ratios of radioactive isotopes among all fractions whereby at least one fraction with deviated ratio of radioactive isotopes is identified as a result said difference is isolated. - View Dependent Claims (7, 8, 9, 10, 11, 12, 13)
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14. A method for finding a difference between two protein samples comprising the steps of:
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(a) labeling protein molecules in each sample with a unique radioactive isotope whereby one isotope is used per sample; (b) mixing the labeled protein molecules from said two samples together into a mixture; (c) contacting said mixture with an array; (d) measuring a ratio of radioactive isotopes at each spot on said array; and
,(e) comparing the ratios of radioactive isotopes among a plurality of spots on said array whereby a spot with deviated ratio of radioactive isotopes can be identified. - View Dependent Claims (15, 16, 17)
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Current AssigneeProteomyx Incorporated
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Original AssigneeProteomyx Incorporated
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InventorsTran, Nathaniel Tue
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Primary Examiner(s)WHISENANT, ETHAN C
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Application NumberUS10/680,277Publication NumberTime in Patent Office924 DaysField of Search435/6, 435/7.1, 530/350, 536/23.1, 536/24.3US Class Current435/6.11CPC Class CodesG01N 2030/77 detecting radioactive prope...G01N 2458/15 Non-radioactive isotope lab...G01N 33/60 involving radioactive label...G01N 33/6848 Methods of protein analysis...
