Epidermal Growth Factor Receptor Tyrosine Kinase Assays in Single Intact Cells Using Capillary Electrophoresis Public Deposited

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  • March 19, 2019
Creator
  • Turner, Abigail
    • Affiliation: College of Arts and Sciences, Department of Chemistry
Abstract
  • Quantification of abnormal Epidermal Growth Factor Receptor (EGFR) tyrosine kinase activity is critical to the clinical success of targeted inhibitors used to treat EGFR-dependent cancers. Current selection criteria based on the presence or absence of activating mutations in the EGFR gene need to be supplemented with complementary assays which quantify EGFR biochemical activity in intact living cells. While in cellulo EGFR activity can be measured to great success in tissue cultured cells using genetically encoded reporters, such techniques are incompatible with small, highly heterogeneous tumor samples. A chemical cytometry approach to direct EGFR activity measurements is presented which consists of an in cellulo peptide reporter-based kinase assay coupled to ultrasensitive microelectrophoretic analysis. The major challenge to such a measurement is biological assay interference from off-target enzymes. A novel approach to minimizing selectivity-based assay interference and its application in single intact cells is presented.
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  • In Copyright
Advisor
  • Allbritton, Nancy
  • Wightman, R. Mark
  • Lawrence, David
  • Lockett, Matthew
  • Der, Channing
Degree
  • Doctor of Philosophy
Degree granting institution
  • University of North Carolina at Chapel Hill Graduate School
Graduation year
  • 2015
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  • Chapel Hill, NC
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