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Protein kinases regulate nearly every aspect of cell life. They participate in many important physiological processes and signalling pathways are closely associated with phosphorylation of proteins and lipids. In addition, alterations in expression and mutations in gene expression cause human diseases, predominantly cancer, but also non-cancerous diseases such as diabetes and hypertension.

Whether you require biochemical or cell based kinase assays, our scientists have long standing expertise in developing both of these for drug discovery.

Kinase Assay Development

We approach kinase assay development by evaluating compounds against a panel of kinases in both biochemical and cellular environments. 

This expertise is coupled with proactive communication by our experienced scientists. 

As a result our team enable you to understand the kinetic profile of your compounds and how they behave in a physiological environment.

Below we outline the advantages and disadvantages of different assays.

Kinase Assays

Biophysical assay (SPR, ITC etc) 

These are used to examine the binding of a compound to the kinase protein.


    • Provides binding kinetics, including on and off rates


    • Only the binding pocket, not the complete kinase is used due to problems in expression and purification of full-length protein
    • Throughput can be limited if compounds have slow off-rates, consequently this will reduce the rate that the biophysical binding surface can be regenerated

Biochemical assay

To measure the turnover of ADP to ATP generally, through a coupled reaction that generates a measurable product (e.g. ADP Glo – Promega).


    • Applicable to all kinase if co-factors etc are known and provided for the binding
    • Provides turnover information (rate of reaction)


    • Does not examine cellular permeability
    • Can be difficult to develop if the ATP turnover is slow

Cell-based target engagement assay

Examines the competitive displacement of a pan-binding tracer (small molecule tagged with a fluorophore) to the kinase in the presence of competing compounds. View our list of target engagement kinase assays here


    • Full length kinase expressed
    • Competition assay occurs within cells under physiological ATP and pH
    • Gives cell penetration data of compounds


    • A small molecule is required to act as the tracer and requires a fluorophore to be added and therefore adds cost and time and may change the binding properties

Our Track Record

View the following examples of projects carried out in this arena and how we supported:

  • Carrick Therapeutics in the development of CT7001 – a CDK-7 inhibitor for breast cancer
  • Nflection to deliver the first clinical trial of NFX-179 Gel – a MEK inhibitor that decreases phopspho-ERK and is used as a topically applied therapy for Cutaneous Neurofibromatosis Type I.

FAQs of Kinases & Kinase Assays

We take a look at some of the most common questions regarding kinases and offer answers on what they are, their uses and structure.

A kinase is an enzyme that catalyses the transfer of phosphate groups from high energy, phosphate-donating molecules to specific substrates, a process known as phosphorylation.

In this process, high energy ATP molecules donate a phosphate group to the substrate molecule generating a phosphorylated substrate and ADP.

Conversely, proteins can donate the phosphate group to ADP (dephosphorylation process) to generate ATP and dephosphorylated proteins.

Kinases are used extensively to transmit signals and regulate complex processes in cells. Phosphorylation of signal molecules can enhance or inhibit their activity and modulate their ability to interact with other molecules. Therefore, the addition or removal of phosphoryl groups provides the cell with a means of control. This is because various kinases can respond to different conditions or signals.

Reversible phosphorylation reactions affect and regulate various functional cell processes by regulating and balancing the activity of substrates. Examples include gene expression, cell metabolism, protein synthesis, signal factor release, morphological changes, and apoptosis.

Kinases can also phosphorylate themselves, a process called autophosphorylation. Autophosphorylation is the kinase-catalysed phospho-transfer to a Serine, Threonine or Tyrosine residue on the kinase itself.

Many kinases require autophosphorylation for full or increased catalytic activity for phospho-transfer to other proteins.

The general structure of kinases includes a catalytic domain, a helical domain and multiple binding domains.

Because the role of the kinase is phosphorylation, consequently the catalytic domain is highly conserved, and the three-dimensional structure of kinases are very similar (see diagram below).

The catalytic domain is composed of a bilobate region linked to a hinge region, a G- (or P-) loop region, a catalytic loop, and an activation loop (A-segment). The catalytic cleft between the bilobate region is an ATP binding pocket and is also the region where most ATP-competitive kinase inhibitors bind.

Autophosphorylation occurs on a kinase’s activation loop, in either a trans- or cis- configuration (catalysing phospho-transfer to another kinase molecule or to itself). In addition, an auto-phosphorylated kinase may undergo a conformational change that better suits the kinase to accept exogenous substrates.

Kinase Structure

Example kinase structure

Kinase assays are always under development, so please contact us to discuss your kinase of interest.

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