Direct Evaluation of Akt Kinase Activity with Single Cell Resolution: Innovations in the Development of Peptide Reporters for Use with Capillary Electrophoresis Public Deposited

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  • March 20, 2019
  • Mainz, Emilie
    • Affiliation: College of Arts and Sciences, Department of Chemistry
  • Direct quantification of kinase activity at the single cell level has long been a goal from both a drug development and diagnostic standpoint. This is true for measurements of the protein kinase Akt, which is known to drive aberrant cell growth and metabolism in diseases including rheumatoid arthritis and pancreatic cancer. A more complete mechanistic profile of abnormal Akt activity in these diseases could drive the development of targeted inhibitors or companion diagnostics, but is hindered by the small and heterogeneous samples that can often be obtained from patients. Established tools including genetically encoded sensors and high throughput mass cytometry are incompatible with these primary samples. Herein, a novel chemical cytometry approach is presented which utilizes peptide-based reporters to deliver direct measurements of Akt activity in single cells from cell lines as well as clinical samples. Methodological and chemical innovations resulted in the development of rapid and low cost assay development strategies, use of non-native amino acids as methods to impart additional stability on peptide probes, analysis of individual cells from rheumatoid arthritis subjects, and significant improvement in the overall throughput of analysis.
Date of publication
Resource type
Rights statement
  • In Copyright
  • Tarrant, Teresa
  • Lockett, Matthew
  • Lawrence, David
  • Jorgenson, James
  • Allbritton, Nancy
  • Doctor of Philosophy
Degree granting institution
  • University of North Carolina at Chapel Hill Graduate School
Graduation year
  • 2016

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