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US 10,208,357 B2Granted
Cell based assay
AssigneeMedical Diagnostic Laboratories, LLC
InventorYick Loi Raymond Yu
Life of this patent
Estimating the term…
Abstract
The present invention relates to cell-based assays involving HER2. The assays use assay cells that are transfected with cassettes containing the HER2 gene of interest and measure the effect of mutations on the activity of HER2, and on their response to inhibitors.
Claims16
A method of determining whether an HER2 variant is sensitive to treatment with a HER2 inhibitor in a cell, comprising the steps of:
- a) preparing a cDNA encoding said HER2 variant;
- b) preparing an expression cassette containing the HER2 variant cDNA;
- c) transfecting said prepared expression cassette containing said HER2 variant cDNA in an assay cell having a JNK reporter construct having SEQ ID NO:
- 31 comprising a reporter gene cDNA linked to at least one AP-1 binding site, and said cell is capable of expressing HER3, wherein the HER2 variant and HER3 are expressed;
- d) exposing said transfected cell to a HER3 ligand, wherein HER3 complexes with said HER2 variant to form a dimer which thereby activates the JNK reporter construct and generates a signal;
- e) exposing said transfected cell with an HER2 inhibitor;
- and
- f) determining whether said HER2 variant is sensitive to treatment with said HER2 inhibitor by measuring a change in signal.
5 more elements
A method of determining whether an HER2 variant is sensitive to treatment with an HER2 inhibitor in a cell, comprising the steps of:
- a) preparing a cDNA encoding said HER2 variant;
- b) preparing an expression cassette containing the HER2 variant cDNA;
- c) transfecting said prepared expression cassette containing said HER2 variant cDNA in an assay cell having a stably integrated JNK reporter construct having SEQ ID NO:
- 31 comprising a reporter gene cDNA linked to 6 AP-1 binding sites, said assay cell having a stably integrated HER3 expression construct;
- and said assay cell having a HER1 and HER2 double knockout, wherein the HER2 variant and HER3 are expressed;
- d) exposing said transfected cell to a HER activator, wherein HER3 complexes with said HER2 variant to form a dimer which thereby activates the JNK reporter construct and generates a signal;
- e) exposing said transfected cell with an HER2 inhibitor;
- and
- f) determining whether said HER2 variant is sensitive to treatment with said HER2 inhibitor by measuring a change in signal.
6 more elements
Dependencies are inferred from the claim text.