Method Development of Chemical Isotope Labeling of Cellular Extract and Biofluids on Metabolomics Profiling and Its Application for Disease Biomarker Discovery

• Author / Creator

  Gu,Xinyun

• Metabolomics aims at studying all the small molecules in biological samples. Compared to transcriptome and proteome, metabolome is highly sensitive to diverse individuals and environmental factors. Thus, metabolomics study has been used to understand individual variations caused by genes, exosomes, diseases, and other metabolic activity and became a very useful tool in biomarker discovery. In order to detect and quantify certain biomarkers from the whole metabolome, high coverage profiling and accurate quantification are essential. Conventional approaches combined several complementary methods to improve low coverage. Our chemical isotope labeling (CIL) LC-MS stands out because of a simplified platform and an overall promoted analytical performance of metabolites.
  My research focuses on utilizing CIL LS-MS methods to profile an amine/phenol submetabolome to study the impact of diseases on biofluids such as cell extracts and serum samples. In the first part of my thesis, we used CIL LC-MS to evaluate and compare two cell harvest methods (physical scraping and trypsinization) and two cell lysis methods (glass-bead-assisted lysis and freeze-thaw-cycle lysis). In the seconded part, we applied the optimized cell harvest protocol to the cell extract, followed by routine LC-MS analysis. By combining the cell results and serum results, we found several metabolites that could be potential biomarkers for Hyper IgE syndrome. In another biomarker discovery study of metformin treatment of diabetes (Chapter 4), CIL LC-MS was applied to reveal the metabolome differences among control, obese, diabetes, and metformin treated groups.

• Subjects / Keywords

  • Chemical Isotope Labeling
  • Metabolomics

• Graduation date

  Fall 2019

• Type of Item

  Thesis

• Degree

  Master of Science

• DOI

  https://doi.org/10.7939/r3-7mah-k592

• License

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