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US 10,379,126 B2Granted
Systems and methods for proteomic analysis
AssigneeThe Regents of the University of Michigan
InventorHaojie Zhu
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Abstract
Provided herein are compositions and methods for proteomic analysis. In particular, provided herein are compositions and methods for performing mass spectrometry-based protein quantitation analysis.
Claims16
A method of quantitating protein levels, comprising:
- a) obtaining a cell lysate sample comprising i) a plurality of polypeptides;
- and ii) a plurality of full-length stable isotope-labeled polypeptides, wherein said full-length isotope polypeptides are obtained by culturing a cell of the same cell type as said cell lysate sample in stable heavy isotope medium;
- b) extracting and digesting said sample to yield a prepared protein fraction;
- c) analyzing said prepared protein fraction with a liquid chromatography-mass spectrometry/mass spectrometry system to obtained mass spectrometry peak areas for said plurality of polypeptides and said full length isotope-labeled polypeptides;
- d) calculating relative peak areas of said plurality of polypeptides and said full length isotope-labeled polypeptides to obtained relative quantitative values;
- and
- e) comparing the relative peak area ratio of said polypeptides to a standard curve generated using protein quantitation standards to determine the absolute quantity of said plurality of polypeptides.
4 more elements
Dependencies are inferred from the claim text.