Receptor tyrosine kinase assays

US8067155B2 · kind B2 · utility

Assignee

• DISCOVERX CORPORATION · US

Inventors

• Wei Feng · Little Rock, US
• William Raab · San Francisco, US
• Philip Achacoso · Union City, US
• Thomas S. Wehrman · Redwood City, US
• Keith R. Olson · Pleasanton, US

Key dates

• Filing date: Aug 6, 2009
• Grant date: Nov 29, 2011
• Expiry date: Aug 6, 2029

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/485
• WIPO field: Measurement
• WIPO sector: Instruments

Abstract

Methods for detecting phosphorylation of receptor tyrosine kinases (“RTKs”) upon activation are provided. The method employs cells comprising two fusion products: (1) an RTK fused to a small fragment of β-galactosidase and (2) a phosphotyrosine binding peptide fused to the large fragment of β-galactosidase, where the 2 fragments weakly complex to form an active enzyme, and optionally a construct for a cytosolic RTK phosphorylating kinase, when the RTK does not autophosphoryate. To detect phosphorylation a β-galactosidase substrate is added to the cells, whereby product formation indicates the occurrence of phosphorylation.