In Vitro Efficacy Testing Services for Primary Biliary Cholangitis

We provide robust and sensitive in vitro screening and characterization platforms for accelerating the discovery and screening of potential therapies for Primary Biliary Cholangitis (PBC). Our service offers comprehensive assays to evaluate compound efficacy, mechanism of action, and pathway modulation specifically relevant to PBC. Key targets in this disease include nuclear receptors such as FXR and PPARs, immune signaling proteins, and cholangiocyte function pathways. Our platforms enable assessment of bile acid metabolism, inflammatory signaling, and fibrogenic processes associated with the pathological progression of PBC.

Our testing portfolio encompasses a wide range of biochemical, cell-based, and reporter assays designed to evaluate drug-target interactions, functional activity, and cellular responses. These methods enable assessment of ligand binding, receptor activation, gene transcription, protein recruitment, and pathway modulation. Our integrated approach supports comprehensive profiling of candidate compounds for efficacy and mechanism of action.

Arrestin protease recruitment assay: Measures the recruitment of arrestin to activated receptors, providing insight into receptor signaling and desensitization relevant to cholangiocyte function.

Cell-based assay: Evaluates compound effects on live cells, supporting the study of cellular toxicity, proliferation, and pathway-specific responses in cholangiocytes.

Chemiluminescent assay: Detects target activity or biomarker expression via light emission, offering high sensitivity for signaling pathway analysis.

Competitive binding assay: Assesses the ability of compounds to compete with labeled ligands for receptor binding, determining binding affinity and specificity.

Displacement of [3H]-dexamethasone: Measures the ability of test compounds to displace radiolabeled dexamethasone from its receptor, useful for profiling glucocorticoid receptor interactions.

Displacement of [3H]-farglitazar: Evaluates the displacement of radiolabeled farglitazar, supporting PPAR receptor binding studies relevant to metabolic regulation in PBC.

ELISA assay: Quantifies specific proteins or biomarkers in cell supernatants or lysates, facilitating the measurement of inflammatory and fibrotic mediators.

Fluorescence resonance energy transfer (FRET) assay: Monitors molecular interactions through energy transfer between fluorophores, useful for detecting protein-protein or ligand-receptor interactions.

Fluorescent assay: Uses fluorescence-based readouts to detect changes in cellular activity, target binding, or reporter expression.

Fluorescent polarization assay: Measures binding interactions based on changes in fluorescence polarization, providing affinity data for drug-target interactions.

Gene reporter assay: Detects gene transcriptional activity by measuring reporter gene expression, enabling functional analysis of pathway modulation.

Homogeneous Time Resolved Fluorescence (HTRF) assay: Combines FRET and time-resolved measurement for sensitive detection of biomolecular interactions in a homogeneous format.

Luciferine/luciferase assay: Quantifies cellular activity or gene expression by measuring bioluminescence, often used in transcriptional activity assays.

NCOR1 (NR box1) peptide recruitment assay: Assesses recruitment of nuclear receptor co-repressor peptides, informing on ligand-induced receptor conformation and activity.

Nuclear translocation assay: Monitors the movement of proteins or receptors into the nucleus, indicating activation of transcriptional pathways.

Peroxisome beta-oxidation assay: Measures fatty acid oxidation activity within peroxisomes, relevant for assessing metabolic pathway modulation in PBC.

RNA assay: Quantifies gene expression changes at the RNA level, supporting downstream pathway and biomarker analysis.

Radioactivity assay: Detects radiolabeled ligand or substrate binding and turnover, providing quantitative data on receptor occupancy and activity.

SRC-1 peptide recruitment assay: Evaluates the interaction of steroid receptor coactivator-1 peptides with nuclear receptors, supporting assessment of receptor activation.

SRC-1-2 peptide recruitment assay: Similar to SRC-1 assay, tests recruitment of a different region of the coactivator for comprehensive profiling.

SRC-2-2 peptide recruitment assay: Measures recruitment of another coactivator peptide to nuclear receptors, enabling mechanistic studies.

Surface plasmon resonance assay: Provides real-time, label-free measurement of binding kinetics and affinities between biomolecules.

Transactivation assay: Assesses the ability of compounds to activate nuclear receptors and induce gene transcription, critical for evaluating functional efficacy.

Transcription assay: Measures overall gene transcriptional activity, allowing assessment of pathway activation or inhibition.

Translocation assay: Detects movement of proteins between cellular compartments, indicating signaling pathway activation.

beta-Lactamase assay: Reporter assay using beta-lactamase enzyme activity to indicate gene activation, providing sensitive readouts for pathway engagement.

cAMP as substrate: Measures changes in intracellular cAMP levels to assess GPCR or other signaling pathway activity relevant to disease mechanisms.

Our assays deliver a range of quantitative pharmacological parameters, including measures of potency, efficacy, and binding affinity. These parameters are essential for ranking candidate compounds, optimizing lead molecules, and guiding dose selection in preclinical development. Accurate measurement of these values informs decision-making at every stage of the drug discovery process.

EC-50: The concentration of a compound that produces 50% of its maximal effect, indicating potency in functional assays.

IC-50: The concentration of inhibitor required to reduce a specific biological or biochemical function by 50%, used to determine inhibitory potency.

Kd: The equilibrium dissociation constant representing binding affinity between ligand and target; lower values indicate higher affinity.

Ki: The inhibition constant reflecting the binding affinity of an inhibitor for its target, crucial for characterizing competitive inhibitors.

MEC: Minimum effective concentration, the lowest concentration at which a compound shows a desired biological effect.

MED: Minimum effective dose, the smallest dose that achieves the intended therapeutic effect in vitro.

MIC: Minimum inhibitory concentration, the lowest concentration that inhibits visible growth or activity of a target, relevant for antimicrobial screening.

pEC-50: The negative logarithm of the EC-50 value, providing a logarithmic measure of compound potency and facilitating data comparison.

pIC-50: The negative logarithm of the IC-50 value, enabling easier comparison of inhibitor potencies across different assays.

Recommended In Vitro Efficacy Tests

Apolipoprotein A1

Apolipoprotein A1 (ApoA1) is a key HDL component reduced in Primary Biliary Cholangitis (PBC), reflecting disease severity and cholestasis. ApoA1 testing supports PBC drug development by enabling patient stratification and monitoring therapeutic efficacy. Key methods include immunoturbidimetric assays and ELISA. Main parameters measured are serum ApoA1 concentration and its longitudinal changes in response to intervention.

C-X-C Motif Chemokine Ligand 10

C-X-C Motif Chemokine Ligand 10 (CXCL10) is elevated in Primary Biliary Cholangitis (PBC) and contributes to liver inflammation by recruiting immune cells. CXCL10 testing is crucial for evaluating disease activity and therapeutic response in PBC drug development. The service utilizes sensitive ELISA assays to quantify CXCL10 levels, with minimum inhibitory concentration (MIC) as a key parameter for assessing drug efficacy in modulating CXCL10-mediated pathways.

Cytochrome P450 Family 7 Subfamily A Member 1

Cytochrome P450 Family 7 Subfamily A Member 1 (CYP7A1) regulates bile acid synthesis, playing a critical role in Primary Biliary Cholangitis (PBC) pathogenesis. Testing CYP7A1 expression via RNA assays enables assessment of drug effects on bile acid metabolism, supporting PBC drug development. The main parameter, Minimal Effective Dose (MED), guides dose optimization for therapeutic efficacy while minimizing adverse effects. This service ensures targeted, mechanism-based evaluation for novel PBC therapies.

Nuclear Receptor Subfamily 1 Group H Member 4

Nuclear Receptor Subfamily 1 Group H Member 4 (NR1H4/FXR) regulates bile acid homeostasis and is pivotal in Primary Biliary Cholangitis (PBC) pathogenesis. Testing NR1H4 activity is crucial for developing targeted PBC therapies. We offer advanced cell-based assays, FRET, chemiluminescent, fluorescent, HTRF, peptide recruitment, beta-lactamase, luciferase, transactivation, and gene reporter assays to assess compound efficacy. Key parameters include EC-50, pIC-50, MEC, pEC-50, IC-50, and Kd.

Nuclear Receptor Subfamily 3 Group C Member 1

Nuclear Receptor Subfamily 3 Group C Member 1 (NR3C1/Glucocorticoid Receptor) modulates immune responses implicated in Primary Biliary Cholangitis (PBC) pathogenesis. Testing NR3C1 is critical for PBC drug development to assess compound efficacy and selectivity. We offer FRET, chemiluminescent, fluorescent, HTRF, nuclear translocation, luciferase, gene reporter, and [3H]-dexamethasone displacement assays. Key parameters—EC-50, pEC-50, IC-50, and Ki—are measured to guide therapeutic optimization.

Peroxisome Proliferator Activated Receptor Alpha

Peroxisome Proliferator Activated Receptor Alpha (PPARα) regulates lipid metabolism and inflammation, both central to Primary Biliary Cholangitis (PBC) pathology. PPARα testing is vital for developing targeted PBC therapeutics. Our service employs advanced assays (e.g., FRET, chemiluminescence, HTRF, transactivation, and gene reporter) to assess ligand affinity, receptor activation, and transcriptional effects. Key parameters measured include EC-50, MEC, IC-50, Kd, and Ki, ensuring robust drug candidate characterization.

Peroxisome Proliferator Activated Receptor Delta

Peroxisome Proliferator Activated Receptor Delta (PPARδ) plays a crucial role in modulating inflammation and bile acid metabolism in Primary Biliary Cholangitis (PBC). Testing PPARδ activity is vital for identifying and optimizing drug candidates for PBC. Using advanced assays—including FRET, HTRF, chemiluminescent, luciferase, gene reporter, and surface plasmon resonance—key pharmacological parameters such as EC50, IC50, Kd, and Ki can be precisely measured to guide drug development.

Peroxisome Proliferator Activated Receptor Gamma

Peroxisome Proliferator Activated Receptor Gamma (PPARγ) plays a crucial role in modulating inflammation and fibrosis in Primary Biliary Cholangitis (PBC). Testing PPARγ activity is vital for screening and optimizing drug candidates targeting this pathway. Key methods include FRET, chemiluminescent, HTRF, competitive binding, luciferase, gene reporter, and surface plasmon resonance assays. Main parameters assessed are EC-50, MEC, IC-50, Kd, and Ki for evaluating drug efficacy and binding affinity.

Phosphodiesterase 4B

Phosphodiesterase 4B (PDE4B) regulates cAMP levels, contributing to inflammation in Primary Biliary Cholangitis (PBC). Testing PDE4B inhibitors is crucial for PBC drug development. Key methods include fluorescent polarization and chemiluminescent assays using cAMP as a substrate, alongside RNA assays for expression analysis. Main parameters measured are MIC and IC-50, providing vital data on inhibitory potency and efficacy of candidate compounds.

Sphingosine-1-Phosphate Receptor 4

Sphingosine-1-Phosphate Receptor 4 (S1PR4) modulates immune cell function, contributing to inflammation in Primary Biliary Cholangitis (PBC). Testing S1PR4 activity is crucial for identifying and optimizing targeted therapies. Our service uses a sensitive arrestin protease recruitment assay to evaluate compound effects on S1PR4, providing precise EC-50 measurements. This enables effective screening and characterization of drug candidates for PBC treatment development.

Sphingosine-1-Phosphate Receptor 5

The Sphingosine-1-Phosphate Receptor 5 (S1PR5) is implicated in immune cell regulation in Primary Biliary Cholangitis (PBC). Testing S1PR5 activity aids in evaluating potential PBC drug candidates. Our service utilizes the Arrestin protease recruitment assay to quantify receptor activation, providing precise EC-50 values. This enables accurate assessment of drug potency and efficacy, supporting informed decisions in PBC drug development.

Superoxide Dismutase 1

Superoxide Dismutase 1 (SOD1) is implicated in oxidative stress modulation in Primary Biliary Cholangitis (PBC), a key factor in disease progression. SOD1 testing is crucial for evaluating oxidative injury and therapeutic response in PBC drug development. Key methods include enzymatic activity assays and immunodetection (ELISA, Western blot). Main parameters measured are SOD1 expression levels and enzymatic activity, providing vital biomarkers for drug efficacy and safety assessment.