Protein Kinase Selectivity Profiling Using Microfluid Mobility Shift Assays
Biochemical selectivity profiling is an integral part of early drug development. Typically compounds from optimization phase are regularly tested for off-target activities within or across target families. This article presents workflow and critical aspects of biochemical protein kinase profiling based on microfluidic mobility shift assays.
Key wordsKinase Selectivity profiling Microfluidic mobility shift assay Compound preparation
I would like to thank Shin Numao and Patrik Roethlisberger for their helpful input to the manuscript. I would like to thank Joerg Trappe for suggesting and encouraging the drafting of this manuscript.
- 1.Fischer EH, Krebs EG (1955) Conversion of phosphorylase b to phosphorylase a in muscle extracts. J Biol Chem 216:121–132Google Scholar
- 2.Krebs EG, Kent AB, Fischer EH (1958) The muscle phosphorylase b kinase reaction. J Biol Chem 231:73–83Google Scholar
- 6.Comley J (2013) Outsourced kinase profiling services - adding value to in-house kinase programmes. Drug Discov World Fall 2013:26–45Google Scholar
- 8.Li H (2009) Review of biochemical assays for protein kinase drug discovery. Trends Bio/Pharmaceutical Ind 5(1):24–32Google Scholar
- 10.Heedmann B, Klumpp M (2015) Screening for inhibitors of kinase autophosphorylation. In: Janzen WP (ed) High throughput screening: methods and protocols, 3rd edn. Springer, New York, NYGoogle Scholar