List of the InVEST Panel (36 targets)
|Target family||Target name||HGNC reference||Additional synonyms||Assay format||Species|
|Cytochrome P450||CYP1A2||CYP1A2||CYP 1A2||Enzymatic activity||Human|
|Cytochrome P450||CYP2C19||CYP2C19||CYP 2C19||Enzymatic activity||Human|
|Cytochrome P450||CYP2C9||CYP2C9||CYP 2C9||Enzymatic activity||Human|
|Cytochrome P450||CYP2D6||CYP2D6||CYP 2D6||Enzymatic activity||Human|
|Cytochrome P450||CYP3A4||CYP3A4||CYP 3A4||Enzymatic activity||Human|
|Cytochrome P450||CYP19A||CYP19A||Aromatase, CYP19A1, CYP 19A||Enzymatic activity||Human|
|GPCR||Serotonin 5-HT2A Receptor||HTR2A||5-hydroxytryptamine receptor 2A, 5-HT-2A, Serotonin receptor 2A||Radioligand filter binding||Human|
|GPCR||Adenosine A2A Receptor||ADORA2A||ADORA, ADORA2, RDC8, Adenosine receptor A2a||Radioligand filter binding||Human|
|GPCR||Opioid kappa Receptor||OPRK1||opioid receptor, Kappa-type opioid receptor kappa 1||Radioligand filter binding||Human|
|GPCR||Opioid mu Receptor||OPRM1||opioid receptor, Mu-type opioid receptor mu 1||Radioligand filter binding||Human|
|GPCR||Serotonin 5-HT2B Receptor||HTR2B||5-hydroxytryptamine receptor 2B, 5-HT2B, Serotonin receptor 2B||Radioligand filter binding||Human|
|GPCR||Serotonin 5-HT1B Receptor||HTR1B||5-hydroxytryptamine receptor 1B, 5-HT1B, Serotonin receptor 1B||Radioligand filter binding||Human|
|GPCR||Muscarinic M3 Receptor||CHRM3||Cholinergic receptor muscarinic 3, Muscarinic acetylcholine receptor M3||Radioligand filter binding||Human|
|GPCR||Serotonin 5-HT1A Receptor||HTR1A||5-hydroxytryptamine receptor 1A, 5-HT1A, Serotonin receptor 1A||Radioligand filter binding||Human|
|GPCR||Muscarinic M2 Receptor||CHRM2||Cholinergic receptor muscarinic 2, Muscarinic acetylcholine receptor M2||Radioligand filter binding||Human|
|GPCR||Adenosine A1 Receptor||ADORA1||RDC7, Adenosine receptor A1||Radioligand filter binding||Human|
|GPCR||Dopamine D3 Receptor||DRD3||D(3) dopamine receptor||Radioligand filter binding||Human|
|GPCR||Opioid delta Receptor||OPRD1||opioid receptor, Delta-type opioid receptor delta 1||Radioligand filter binding||Human|
|GPCR||Dopamine D2S Receptor||DRD2||D(2) dopamine receptor||Radioligand filter binding||Human|
|GPCR||Histamine H1 Receptor||HRH1||Histamine H1 receptor, HH1R, H1R||Radioligand filter binding||Human|
|GPCR||Adrenergic Alpha 2A Receptor||ADRA2A||ADRAR, ADRA2, ADRA2R,Alpha-2A adrenergic receptor, adrenoceptor alpha 2A||Radioligand filter binding||Human|
|GPCR||Adrenergic Alpha 1A Receptor||ADRA1A||Alpha-1A adrenergic receptor, adrenoceptor alpha 1A||Radioligand filter binding||Rat|
|GPCR||CCK1 Receptor||CCL28||SCYA28, MEC, CC chemokine CCL28, C-C motif chemokine 28||Radioligand filter binding||Human|
|GPCR||Muscarinic M1 Receptor||CHRM1||Cholinergic receptor muscarinic 1, Muscarinic acetylcholine receptor M1||Radioligand filter binding||Human|
|GPCR||Dopamine D1 Receptor||DRD1||D(1A) dopamine receptor||Radioligand filter binding||Human|
|Ion Channel||hERG||KCNH2||potassium voltage-gated channel subfamily H member 2, Ether-a-go-go-related gene potassium channel 1, Kv11.1||Fluorescence Polarization||Human|
|Ion Channel||Serotonin 5-HT3 Receptor||HTR3A||5-hydroxytryptamine receptor 3A, 5-HT3A, Serotonin-gated ion channel receptor, HTR3||Radioligand filter binding||Human|
|Ion Channel||GABA-A Receptor||GABRA1||gamma-aminobutyric acid type A receptor, GABA(A) receptor||Radioligand filter binding||Rat|
|Nuclear Receptor||Glucocorticoid receptor||NR3C1||GR, GRL, nuclear receptor subfamily 3 group C member 1||Fluorescence Polarization||Human|
|Nuclear Receptor||Estrogen receptorβ||ESRRB||ERR-beta, ERRB2, Steroid hormone receptor ERR2, ERR beta-2, Estrogen receptor-like 2, NR3B2||Fluorescence Polarization||Human|
|Nuclear Receptor||Progesterone Receptor||PGR||PR, NR3C3, Nuclear receptor subfamily 3 group C member 3||Fluorescence Polarization||Human|
|Nuclear Receptor||Aryl Hydrocarbon Receptor||AHR||Ah receptor, BHLHE76||Cell-based transcriptional regulation||Human|
|Phosphodiesterase||PDE3A||PDE3A||cGMP-inhibited 3',5'-cyclic phosphodiesterase A||Enzymatic activity||Human|
|Phosphodiesterase||PDE4A||PDE4A||cAMP-specific 3',5'-cyclic phosphodiesterase 4A||Enzymatic activity||Human|
|Protease||Thrombin||F2||coagulation factor II||Enzymatic activity||Human|
We consistently establish more targets for our InVEST Panel. Please reach out to inquire if the target of your interest is currently in development.
|Category||Target name||HGNC reference||Synonyms||Data|
|Calcium Ion Channel||Cav1.2 Ion Channel||CACNA1C||Cav1.2, CACH2, CACN2, TS, LQT8, Voltage-dependent L-type calcium channel subunit alpha-1C||Data sheet|
|Potassium Ion Channel||hERG Ion Channel||KCNH2||potassium voltage-gated channel subfamily H member 2, Ether-a-go-go-related gene potassium channel 1, Kv11.1||Data sheet|
|Sodium Ion Channel||Nav1.5 Ion Channel||SCN5A||sodium voltage-gated channel alpha subunit 5, Nav1.5 LQT3, HB1, HBBD, PFHB1, IVF, HB2 HH1 SSS1 CDCD2, CMPD2, ICCD||Data sheet|
Additonal information to InVEST Panel Screening
Setups: Standard panel testing consists of single concentration testing in duplicates. Alternate testing formats may be available upon request.
Binding assays: Total binding (DMSO vehicle) and non-specific binding (containing saturating concentration of reference compound) and a full concentration-response curve with an appropriate reference compound are conducted with each target.
Enzymatic activity assays: No inhibitor control (DMSO vehicle) and a full concentration-response curve with an appropriate reference compound are conducted with each target.
Turnaround time: 10 business days for standard projects. Expedited scheduling and data delivery can be arranged prior to the commencement of the studies.
Report: The raw data, % inhibition, 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.
Screening facility: This assay is performed at our screening facility in Malvern, PA, US.
Compound requirements: In brief, for a standard project with a final test concentration of 10 µM we require 100 µl of a 10 mM stock solution in DMSO or 5 mg powder. Less material per test is needed for large-scale screening. Please refer to our FAQs for information regarding compound preparation and shipping.
Radioligand Binding Assay
A. Commercially sourced membrane preparations and prepared rat brain membranes expressing the target receptors, respectively, are placed in multi-well plates. B. 3H-labelled ligands incubate with the membrane preparations in the presence and absence of test compounds for 90 minutes. C. The solution is pulled through a filter trapping the membranes and bound ligands. Unbound ligands are not retained on the filter. The radioactivity in the filter is measured via scintillation counting as a readout of ligand-target receptor binding.
The fluorescence polarization (FP) assays are binding assays where a ligand conjugated to a fluorophore binds to the target receptor.
A. The FP assay is performed with either soluble protein or (in the case of hERG) membrane preparations. B. The client compound and the fluorescent ligand incubate with the target. C. Ligand binding is quantified by measuring the amount of polarized light emitted by the fluorophores in the well.
Principle: Light waves, passed through certain crystalline materials (polarizers), have their electrical vectors oriented in a single plane and are thus polarized. The fluorophores linked to the ligand absorb the light and emit fluorescence. Soluble, small fluorescent ligands have a high rotational movement (Brownian movement) that is shorter than their fluorescence decay time resulting in their emitted fluorescence light being depolarized. Fluorescent ligands bound to the larger receptor have a rotational movement that is slower than their fluorescence decay time thus, their emitted light remains polarized.
Cell-based transcriptional regulation
Receptor-specific reporter cells express the luciferase reporter gene functionally linked to a bona fide receptor-responsive promoter. Thus, quantifying changes in luciferase expression in the treated reporter cells provides a sensitive surrogate measure of the changes in receptor activity. This assay format is suitable to investigate agonistic and antagonistic responses of the target receptor.
The next upcoming run starts on April 18th. The compound receiving deadline is April 13th.
Testing of the InVEST Panel is performed on a monthly basis. Please reach out to learn about upcoming screening dates.
Fluorescence Polarization Assay with a Nuclear Receptor
Progesterone Receptor Fluorescence Polarization: Results of three independent experiments of progesterone binding are shown. 95% CV shown as dashed lines. Sigmoid fit (solid line) parameters: IC50=36.9 nM, Hill slope=−2.13. 95% confidence intervals: EC50: 30.4 to 44.8 nM, Hillslope: ‑3.0 to ‑1.2.
Fluorescence Polarization Assay with an Ion Channel
hERG Receptor Fluorescence Polarization: Results of six independent experiments of E-4031 binding to hERG containing membranes are shown.Average of 6 independent experiments. 95% CV shown as dashed lines. Competing drug: E-4031. Sigmoid fit (solid line) parameters: IC50=20.9 nM, Hill slope=−1.46. 95% confidence intervals: IC50: 14.7 to 29.7 nM, Hillslope: ‑-2.08 to ‑0.84.
Radioligand Assay with a GPCR
Histamine H1 Receptor Radioligand Binding Assay: Results of one experiment performed in duplicates of binding of reference compound pyrilamine to Histamine H1 receptor are shown.
Sigmoid fit parameters: EC50=1.25 nM, Hill slope=−0.99.
Enzymatic Activity Assay with a PDE
Three reference compounds IBMX, methoxyquinazoline, and Rolipram were tested against the activity of cAMP-specific cyclic phosphodiesterase 4A (PDE4A). Concentration-response curves are shown with semi-log concentrations in singlicates with the following parameters:
IBMX: IC50= 1.4e-05, hillslope= -0.72
Methoxyquinazoline: IC50= 7.82e-07, hillslope= -0.86
Rolipram: IC50= 1.1e-06, hillslope= -0.86
Enzymatic Activity Assay with a Protease
Reference inhibitor gabexate mesylate (GM) was tested against thrombin for IC50 value determination with semi-log concentrations in singlicate. The fluorescent substrate Pefafluor TH containing AMC was used as a substrate. Parameters: IC50= 5.9e-07, hillslope= -0.94