Kinase Panel Screening and Profiling Service

Broad kinase screening is crucial for the identification of selective and potent kinase inhibitors. Determination and improvement of the selectivity of a compound by screening a large part of the kinome are of pivotal importance in the discovery process of novel kinase inhibitors e.g. to reduce the risk of adverse side effects or both, kinase inhibitors and compounds inhibiting non-kinase targets.

Reaction Biology offers the largest kinase panel for kinase profiling in the industry ensuring the most reliable results for your inhibitor selectivity screening covering most of the human kinome.
Kinase selectivity profiling is performed on a monthly (³³PanQinase™) or bi-weekly (HotSpot™) schedule, which allows us to offer very competitive pricing.
HotSpot kinase screening service now available at physiologically relevant 1mM ATP for 340 wild type kinases, in addition to existing standard concentrations of 1μM, 10μM or apparent ATP-Km up to 100μM
Specialty panels comprise a selection of kinase-specific mutants or our CDK panel.
Kinase panel reports from the US facility include a free control compound’s IC50 curve.

Reaction Biology’s kinase assays were ranked highest in the Kinase Profiling Trends Survey by HTStec for a reason: Our contract research organization accelerates the drug discovery and development process in two global facilities: one in Pennsylvania, one in Germany. We learn about what our clients want to achieve in their research and match it with our most suitable kinase selectivity profiling service.

As our panels of kinases continue to grow, we consistently demonstrate the most reliable kinase selectivity screening services for clients worldwide based on accurate kinase activity data. Contact us today to discuss how our kinase panel assays can aid your project!

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New Poster: Kinase drug discovery 20 years after imatinib

Reaction Biology sponsored a poster authored by Philip Cohen, Darren Cross, and Pasi Jänne that was published by Nature Reviews Drug Discovery.

The peer-reviewed poster illustrates the tremendous progress made over the last 20 years towards the development of new kinase inhibitors.

Download our Poster

RB Kinase Drug Discovery Poster

Selectivity Kinase Panels from Reaction Biology are the Largest in the Industry with 700+ Kinases

Panels run at US facility
Panels run at US facility	Panel size	ATP options	Testing options	Panel
Wild Type Kinase Panel	384	1μM, 10μM, Km and 1mM	1 & 10 doses	view panel
Diversify Panel	69	Km & 1mM	1 & 10 doses	view panel
Mutant Kinase Panel	355	1μM, 10μM, Km	1 & 10 doses	view panel
Atypical Kinase Panel	24	1μM, 10μM, Km	1 & 10 doses	view panel
Lipid Kinase Panel	23	1μM, 10μM, Km	1 & 10 doses	view panel
Diacylglycerol Kinase Panel	10	50μM, 200μM, Km & 1mM	1 & 10 doses	view panel

Kinase Selectivity Assay Panels
Panels run at German facility	Panel size	ATP options	Testing options	Panel
Wild Type Kinase Panel	355	Km	1 & 10 doses	view panel
Mutant Kinase Panel	96	Km	1 & 10 doses	view panel
Lipid Kinase Panel	14	Km	1 & 10 doses	view panel

Specialty Kinase Profiling

In addition to broad kinome selectivity panels, we perform screening of specialty panels on an on-demand basis.

Panels run at US facility	Panel size	Data
ABL1 Mutant Profiler	15	Data sheet
ALK Mutant Profiler	29	Data sheet
BRAF Mutant Profiler	16	Data sheet
c-KIT Mutant Profiler	24	Data sheet
c-MET Mutant Profiler	32	Data sheet
CDK Profiler	35	Data sheet
EGFR Mutant Profiler	65	Data sheet
ERBB2 Mutant Profiler	10	Data sheet
FGFR2 Mutant Profiler	19	Data sheet
FGFR3 Mutant Profiler	6	Data sheet
FGFR4 Mutant Profiler	5	Data sheet
FLT3 Mutant Profiler	13	Data sheet
RET Mutant Profiler	36	Data sheet
TIE Mutant Profiler	10	Data sheet
TRKA Mutant Profiler	15	Data sheet
TRKC Mutant Profiler	6	Data sheet

Panels run at German facility	Panel size	Data
CDK Profiler	32	Data sheet
Lipid Kinase Profiler	14	Data sheet
EGFR Mutant Profiler	11	Data sheet
KIT Mutant Profiler	10	Data sheet
MET Mutant Profiler	10	Data sheet
RET Mutant Profiler	10	Data sheet
ABL1 Mutant Profiler	9	Data sheet
ALK Mutant Profiler	8	Data sheet

Kinase Panel Screening Details

Assay Formats

HotSpot™ and ³³PanQinase™: two radiometric assay formats

Reaction Biology offers two radiometric assay formats that are performed in our screening facilities located in Malvern, PA, USA, and Freiburg, Germany. Both assays are based on the transfer of ³³P-labelled phosphate from ATP to the kinase substrate. The methods, however, differ with respect to retention and detection of the phosphorylated substrates. Proteins and peptides in the HotSpot™ assay (performed in the US) are captured via spotting of the reaction mix on a filter membrane. The ³³PanQinase™ assay (performed in Germany) is performed with ScintiPlate microtiter plates, which are coated with scintillant for detection.

Lipid kinases are screened with ADP-Glo (Promega) at both facilities.

Reproducibility

The IC50 values of a BCR-ABL kinase inhibitor were determined for 320 wild type kinases in two independent experiments and compared. The high R² value shows a high correlation of data. IC50 value determination was performed with 6 compound concentrations in singlicate with the ³³PanQinase™ assay.

The control compound Staurosporine was tested over the course of 6 months in 12 independent experiments. IC50 determination was performed with 10 compound concentrations in singlicate with the HotSpot™ assay.

Assay Setup (US facility)

Screening formats: 2 data point projects with 1 concentration in duplicates and 5 to 10-doses response curves.

Assays: Protein kinases are screened via radiometric HotSpot™ assay. Lipid kinases are screened via ADP-Glo (Promega) assay. The assay protocol for the HotSpot™ assay is available here.

ATP concentrations: 1mM, 1μM, 10μM, or apparent ATP-Km up to 100μM

Controls: No inhibitor (DMSO vehicle) control. For every kinase, we include the screening of one kinase-specific control compound in 10-dose IC50 format.

Timelines: Selectivity panel screening is performed twice per month. Reports are sent 2 to 3 weeks after the project start.

Screening dates: Upcoming selectivity panel screening dates are shown here.

Report: The raw data, % enzyme activity and control compound IC50 values will be reported in Excel format. Assay conditions, target, and substrate information are available upon request. Requirements for this information should be noted prior to the commencement of the study.

Mapper: The results of the selectivity of compounds can be graphically presented with our mapper tool.

Compound requirements: In brief, for kinase panel profiling 100 ul of a 10 mM DMSO stock or solid material is needed. Shipping information can be found in our FAQs.

Please see our FAQ page to access our assay condition form (US facility).

Assay Setup (German facility)

Screening formats: 2 data point projects are performed with 1 concentration in duplicates or 2 concentrations in singlicates. IC50 projects are performed with 6 concentrations in singlicate.

Assays: Protein kinases are screened via radiometric ³³PanQinase™ assay. Lipid kinases are screened via ADP-Glo™ (Promega) assay. The assay protocol for the ³³PanQinase™ assay is available here.

ATP concentrations: apparent ATP-K_m

Controls: DMSO only as full enzymatic activity control.

Timelines: Selectivity panel screening is performed once per month or on-demand. Reports are sent 10 business days after the project start.

Screening dates: Upcoming profiling dates are shown here.

Report: A detailed report will be provided including assay conditions, target, and substrate information. The raw data and % enzyme activity will be reported in Excel format including IC50 values and curve fitting, if applicable. Compound selectivity on wild type protein kinases will be graphically presented with the mapper tool.

Compound requirements: For testing with 1 concentration in duplicate, 1700 µl of a 100x stock solution is required. For testing with 2 concentrations in singlicate, 850 µl of a 100x stock solution of each concentration is required. For IC50 testing, 1000 µl of 100x stock solution is required. Solid material is also accepted.

Please see our FAQ page to access our biochemical assay condition form (German facility) for further details on compound preparation and shipping.

Screening Schedule

Selectivity kinase panels are screened bi-weekly or on a monthly basis

See the screening schedule of the US facility here.

See the screening schedule of the German facility here.

Specialty kinase panels are screened on demand.

Why profiling with IC50 values?

Example of an FLT3 inhibitor with ³³PanQinase™ assay: IC50 values of 320 wild type protein kinases are shown with values of lower than 10^-4. IC50 value determination for selectivity profiling helps assess differences in the potency of a given compound and allows a precise judgment on the selectivity of kinase inhibitors.
Only the targets with the lowest IC50 values are named.

IC50 based profiling is designed to:

Obtain an accurate guiding parameter for optimization of kinase selectivity during the medicinal chemical optimization phase
Ensure that observed functional effects of a kinase inhibitor are not due to the inhibition of off-target-kinases and to identify potential adverse side effects of the drug
Provide accurate comparison to a reference compound

Selectivity and Toxicity

Protein kinases share a common catalytic core and thus usually display sequence and structural similarity, which can result to a lack of selectivity and to off-target toxicity of drug candidates. The lack of selectivity can be beneficial but can also cause adverse toxicities which result in the discontinuation of promising drug candidates.

Book chapter about selectivity and toxicity issues of protein kinase inhibitors