Method for measuring tyrosine kinase phosphorylation

US7981633B2 · kind B2 · utility

Assignee

• Merck Patent GmbH · DE

Inventors

• Tanja Henzler · Schriesheim, DE
• Bjoern Hock · Maintal, DE
• Andree Blaukat · Heilbad Heiligenstadt, DE
• Uwe Hofmann · Würzburg, DE

Key dates

• Filing date: Jul 7, 2006
• Grant date: Jul 19, 2011
• Expiry date: Jul 30, 2026

Classification

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

Abstract

The present invention relates to methods for measuring the autophosphorylation of one or more tyrosine kinases and use of such methods in profiling kinase inhibitors and activators. As a representative example, the method comprises inducing kinase autophosphorylation activity in cells in presence and in absence of a kinase inhibitor, lysing the cells, capturing the tyrosine kinase in the cell lysate by adding a plurality of tyrosine kinase specific binding proteins which are associated with unique dyes, adding a phosphotyrosine specific antibody tagged with a marker which is distinguishable from the unique dyes, and identifying the autophosphorylated tyrosine kinase by detecting the unique dye and the marker. Alternately, the tyrosine kinases themselves could be coupled to the unique dyes. The present invention also relates to kits and compositions for carrying out the above-described methods.