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US 2019/0161787 A1Published application
Methods for predicting kinase inhibitor resistance
AssigneeResearch Development Foundation
InventorsBrent Iverson, George Georgiou, Joseph Desautelle, Joseph Taft
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Abstract
Provided are methods for the identification of mutant kinases that are resistant to inhibition by a kinase inhibitor. In some embodiments, the methods may be used to assess a test compound or kinase inhibitor for the risk of the development of resistance in vivo, e.g., during clinical administration to treat a disease such as a cancer.
Claims47
A method of generating a mutant kinase, comprising:
- (a) expressing in each of a plurality of eukaryotic cells:
- (i) a first fusion protein comprising an ER targeting sequence, a kinase, and an ER retention sequence;
- and
- (ii) a vector encoding a second fusion protein comprising:
- an endoplasmic reticulum (ER) targeting sequence, a surface expression sequence, a first peptide sequence, and a endoplasmic reticulum (ER) retention sequence;
- wherein the eukaryotic cells are exposed to a kinase inhibitor during said expression;
- and
- (b) separating or purifying said eukaryotic cells based on the presence or absence of phosphorylation of at least one amino acid of the first peptide on the surface of the eukaryotic cells.
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