Project Examples – Development of a Kinase Profiler Panel

Objective: To develop a convenient, nonradiometric, and robust assay platform for the measurement of 16 Ser/Thr and 22 Tyr kinases.

Performance Specification: >3-fold signal increase (+)ATP vs. (-)ATP

Proposed Timeline: 16 weeks

Description: IBA evaluated the use of several putative kinase substrates and phospho-specific antibodies in an immunoassay format to achieve the desired signal:noise ratios.

Results: Specific substrates and phospho-antibodies were identified for the detection of 16 Ser/Thr and 22 Tyr kinases.  All assays had a >3-fold signal:noise ratios, utilized the same reaction conditions, and could easily be adapted to both single point and IC₅₀ determinations.  The assay data tightly compared to classical ³²P-ATP assays with several client compounds.

 Project Completion: 12 weeks