In Vitro Efficacy Testing Services for Eosinophilic Esophagitis

We provide robust and sensitive in vitro screening and characterization platforms for accelerating the discovery and screening of potential therapies for Eosinophilic Esophagitis. Our services enable the evaluation of drug candidates targeting key immune pathways, cytokines, and cellular responses implicated in the pathogenesis of this disease. We focus on relevant targets such as eosinophil activation, Th2 cytokine signaling, and epithelial barrier function. Our assays assess processes including inflammation, apoptosis, cellular proliferation, cytokine release, and receptor-ligand interactions to support comprehensive preclinical profiling.

Our in vitro efficacy testing portfolio for Eosinophilic Esophagitis covers a diverse range of biochemical, cellular, and molecular assays. These methods are designed to measure activity, binding affinity, functional impact, and mechanistic action of therapeutic candidates. Collectively, they provide detailed insights into efficacy, potency, and mechanism of action.

ATP assay: Quantifies cellular ATP levels as a measure of cell viability, proliferation, or cytotoxicity.

Annexin V binding assay: Detects early apoptotic cells by binding to phosphatidylserine residues exposed on the cell membrane.

Arrestin protease recruitment assay: Assesses receptor activation by measuring arrestin recruitment, indicative of GPCR signaling.

Biolayer interferometry assay: Measures real-time biomolecular interactions to determine binding kinetics and affinities.

Chemiluminescent assay: Utilizes light emission from chemical reactions to quantify specific analytes or enzymatic activities.

Competitive binding assay: Evaluates the ability of test compounds to compete with labeled ligands for binding to target receptors.

Displacement of [125I]-[Sar1,Ile8]-angiotensin II: Measures the displacement of radiolabeled angiotensin II analogs from receptor binding sites.

Displacement of [125I]-[Tyr4]-angiotensin II: Assesses competition for angiotensin II receptor binding via radioligand displacement.

Displacement of [125I]-angiotensin II: Quantifies compound affinity by measuring the displacement of radiolabeled angiotensin II.

Displacement of [3H]-dexamethasone: Evaluates the ability of compounds to compete for glucocorticoid receptor binding.

Displacement of [3H]-prostaglandin D2: Assesses competitive binding at prostaglandin D2 receptors using tritiated ligand.

Displacement of [3H]-valsartan: Measures receptor-ligand interactions through displacement of radiolabeled valsartan.

Dye assay (alamar blue): Fluorescent dye-based assay for assessing cell viability and metabolic activity.

ELISA assay: Quantitative detection of proteins, cytokines, or antibodies using enzyme-linked immunosorbent techniques.

Flow cytometry assay: Multiparametric analysis of cell populations based on fluorescent labeling of surface or intracellular markers.

Fluorescence resonance energy transfer (FRET) assay: Detects molecular interactions via energy transfer between fluorescent tags.

Fluorescent assay: Utilizes fluorescence-based detection to quantify cellular or molecular events.

Fluorescent-activated cell sorting (FACS) assay: Enables sorting and analysis of specific cell subsets based on fluorescent markers.

Gene reporter assay: Measures gene expression changes by quantifying reporter gene activity under specific promoters.

Homogeneous Time Resolved Fluorescence (HTRF) assay: Non-radioactive, high-sensitivity assay to detect biomolecular interactions or analytes.

Luciferine/luciferase assay: Quantifies gene expression or ATP levels using bioluminescence generated by luciferase enzyme.

Nuclear translocation assay: Measures the movement of signaling molecules or transcription factors into the nucleus.

RNA assay: Detects and quantifies RNA expression levels to assess gene regulation or target engagement.

Radioactivity assay: Employs radiolabeled compounds to quantify binding, uptake, or enzymatic activity.

Saturation binding assay: Determines the maximal binding capacity (Bmax) and affinity (Kd) of compounds for target receptors.

Sensitization with ovalbumin: Models immune sensitization by exposing cells to ovalbumin to assess allergic responses.

Surface plasmon resonance assay: Provides real-time analysis of binding kinetics and affinities between biomolecules.

Thymidine incorporation assay: Measures cell proliferation based on incorporation of radiolabeled thymidine into DNA.

[35S]-GTPgammaS binding assay: Assesses GPCR activation by measuring binding of radiolabeled GTP analogs.

Our assays measure a comprehensive set of pharmacological parameters, including potency, efficacy, affinity, and inhibitory concentrations. These parameters are critical for ranking candidate compounds, optimizing lead selection, and supporting dose-response modeling. Accurate quantification of these metrics enables informed decision-making throughout the drug development process.

EC-300: The concentration of a compound required to achieve 300% of the baseline effect, useful for detailed dose-response analysis.

EC-50: The concentration at which a compound elicits 50% of its maximal effect, indicating potency.

ED-50: The dose required to produce 50% of the maximal therapeutic effect, important for in vitro to in vivo translation.

IC-50: The concentration needed to inhibit a biological process by 50%, commonly used for antagonist evaluation.

Kd: The equilibrium dissociation constant, reflecting the affinity between ligand and receptor.

Ki: The inhibition constant, quantifying the binding affinity of an inhibitor for its target.

MEC: Minimum effective concentration, the lowest concentration at which a compound produces a detectable effect.

MED: Minimum effective dose, the smallest dose achieving a desired therapeutic effect.

MIC: Minimum inhibitory concentration, the lowest concentration inhibiting visible growth of a microorganism or cell population.

pA-2: Negative logarithm of the antagonist dose ratio, used to characterize competitive antagonism.

pEC-50: The negative logarithm of EC-50, providing a more convenient scale for comparing potencies.

pIC-50: The negative logarithm of IC-50, facilitating precise potency comparisons among inhibitors.

pKb: Negative logarithm of the equilibrium constant for antagonist binding, useful for receptor pharmacology.

pKi: Negative logarithm of Ki, commonly used to compare binding affinities across compounds.

Recommended In Vitro Efficacy Tests

Angiotensin Ii Receptor Type 1

The Angiotensin II Receptor Type 1 (AT1R) is implicated in Eosinophilic Esophagitis (EoE) pathogenesis via inflammatory and fibrotic pathways. Testing AT1R is crucial for EoE drug development to assess candidate efficacy and specificity. Our service employs fluorescent and radioactivity assays, including displacement of [125I]- and [3H]-labeled ligands and arrestin recruitment assays. Key parameters measured are pA2, pKi, pIC50, IC50, and Ki, ensuring accurate characterization of AT1R interactions.

Interleukin 13

Interleukin 13 (IL-13) plays a central role in the pathogenesis of Eosinophilic Esophagitis (EoE) by promoting eosinophilic inflammation. Accurate IL-13 testing is crucial for EoE drug development to evaluate therapeutic efficacy and mechanism of action. Key methods include flow cytometry, ELISA, fluorescent, chemiluminescent, surface plasmon resonance, radioactivity assays, and ovalbumin sensitization. Main parameters assessed are ED-50, IC-50, and Kd, providing insights into drug potency and binding affinity.

Interleukin 15

Interleukin 15 (IL-15) is implicated in Eosinophilic Esophagitis (EoE) by promoting eosinophil survival and inflammation. Accurate IL-15 testing is crucial for drug development targeting EoE. We offer chemiluminescent, surface plasmon resonance, and ELISA assays to quantify IL-15 activity and drug interactions. Key parameters measured include IC-50 (inhibitory concentration) and Kd (binding affinity), providing essential data for therapeutic candidate evaluation.

Interleukin 4

Interleukin 4 (IL-4) is a key cytokine driving inflammation in Eosinophilic Esophagitis (EoE). Accurate IL-4 testing is crucial for evaluating drug candidates targeting this pathway. Our service utilizes chemiluminescent assays to quantify IL-4 activity and determine inhibitory concentration 50 (IC-50) values, enabling precise assessment of therapeutic efficacy in EoE drug development.

Janus Kinase 1

Janus Kinase 1 (JAK1) is a key mediator of inflammatory signaling in Eosinophilic Esophagitis (EoE). JAK1 testing is crucial for identifying and optimizing drug candidates targeting this pathway. Our service employs a sensitive fluorescence resonance energy transfer (FRET) assay to assess compound binding affinity, reporting the primary parameter pKi. This enables precise evaluation of potential JAK1 inhibitors for EoE drug development.

Kit Proto-Oncogene, Receptor Tyrosine Kinase

The Kit Proto-Oncogene, Receptor Tyrosine Kinase is implicated in the pathogenesis of Eosinophilic Esophagitis by mediating eosinophil activation and tissue remodeling. Testing its activity is critical for identifying and optimizing targeted therapies. Our service employs chemiluminescent assays to quantify kinase inhibition, providing precise IC-50 values essential for drug candidate evaluation and decision-making in Eosinophilic Esophagitis drug development.

Mechanistic Target Of Rapamycin Kinase

Our Mechanistic Target Of Rapamycin (mTOR) Kinase testing service supports Eosinophilic Esophagitis (EoE) drug development by evaluating mTOR’s key role in regulating immune cell proliferation and inflammation in EoE. This testing identifies compounds that inhibit mTOR, crucial for therapeutic targeting. We employ ATP, ELISA, fluorescent, FRET, chemiluminescent, thymidine incorporation, and RNA assays to determine critical parameters such as IC-50, MEC, Kd, and MIC, ensuring robust candidate profiling.

Nuclear Receptor Subfamily 3 Group C Member 1

Nuclear Receptor Subfamily 3 Group C Member 1 (NR3C1), the glucocorticoid receptor, modulates inflammation in Eosinophilic Esophagitis (EoE). Testing NR3C1 activity is crucial for evaluating drug efficacy and safety in EoE therapy development. Our service employs luciferase, ELISA, fluorescent, FRET, [3H]-dexamethasone displacement, chemiluminescent, HTRF, nuclear translocation, and gene reporter assays. We deliver key pharmacological parameters: MEC, EC-50, MED, EC-300, pEC-50, IC-50, and Ki, supporting robust candidate assessment.

Prostaglandin D2 Receptor 2

Prostaglandin D2 Receptor 2 (DP2/CRTH2) mediates eosinophil activation and recruitment in Eosinophilic Esophagitis, driving inflammation. Testing DP2 is crucial for developing targeted therapies. Key assays include saturation binding, fluorescent, [3H]-prostaglandin D2 displacement, [35S]-GTPγS binding, FACS, and Annexin V binding, enabling assessment of ligand-receptor interactions and cellular responses. Main parameters measured are pA-2, Kd, pKb, IC-50, and Ki, critical for evaluating drug potency and receptor affinity.

Sphingosine-1-Phosphate Receptor 1

The Sphingosine-1-Phosphate Receptor 1 (S1P1) is implicated in Eosinophilic Esophagitis by mediating immune cell trafficking and inflammation. Testing S1P1 activity is crucial for developing targeted therapies. Our service utilizes arrestin protease recruitment and fluorescent assays to assess drug effects on S1P1. Key pharmacological parameters determined include EC50 (potency) and IC50 (inhibition), enabling precise evaluation of candidate compounds for Eosinophilic Esophagitis treatment.

Sphingosine-1-Phosphate Receptor 4

Sphingosine-1-Phosphate Receptor 4 (S1P4) regulates immune cell trafficking and inflammation in Eosinophilic Esophagitis (EoE). Testing S1P4 activity is crucial for identifying potential EoE therapeutics targeting this pathway. Our service utilizes an arrestin protease recruitment assay to measure receptor activation, providing EC-50 values as a key parameter for drug potency and efficacy evaluation. This enables precise candidate selection in EoE drug development.

Sphingosine-1-Phosphate Receptor 5

Our Sphingosine-1-Phosphate Receptor 5 (S1PR5) testing service supports Eosinophilic Esophagitis drug development by assessing S1PR5’s role in modulating immune cell trafficking and inflammation. This testing is crucial for identifying compounds that effectively target S1PR5. Using the Arrestin protease recruitment assay, we precisely measure compound potency, providing key data such as EC-50 values to guide candidate selection and optimization.

Thymic Stromal Lymphopoietin

Thymic Stromal Lymphopoietin (TSLP) is a key cytokine implicated in the pathogenesis of Eosinophilic Esophagitis (EoE) by driving Th2-mediated inflammation. Accurate TSLP testing is vital for EoE drug development to evaluate therapeutic efficacy and mechanism of action. We offer a comprehensive panel of assays—including luciferase, ELISA, FRET, chemiluminescent, flow cytometry, FACS, SPR, BLI, competitive binding, and alamar blue—to quantify TSLP activity and binding, delivering critical parameters such as IC-50 and Kd.