In Vitro Efficacy Testing Services for Pik3Ca Related Overgrowth Spectrum

We provide robust and sensitive in vitro screening and characterization platforms for accelerating the discovery and screening of potential therapies for Pik3Ca Related Overgrowth Spectrum (PROS). Our services are specifically designed to evaluate the efficacy and mechanism of action of candidate compounds targeting the PI3K/AKT/mTOR pathway, which is central to the pathogenesis of PROS. We focus on key molecular targets such as PI3KCA and downstream signaling proteins that drive abnormal cell proliferation and tissue overgrowth. Our assays enable the investigation of therapeutic effects on cellular proliferation, kinase activity, and pathway modulation relevant to the pathological processes underlying PROS.

Our in vitro testing services employ a diverse range of biochemical and cell-based assay technologies tailored to assess compound efficacy, mechanism, and pathway modulation. These methods enable high-throughput and precise evaluation of potential therapeutics' effects on relevant molecular targets and cellular functions. By integrating multiple assay platforms, we provide comprehensive insights into drug activity and candidate prioritization.

ATP assay: Measures cellular ATP levels as a readout of cell viability or metabolic activity, essential for assessing cytotoxicity or proliferation.

Bioluminescence resonance energy transfer (BRET) assay: Enables real-time monitoring of protein-protein interactions or signaling events in live cells, valuable for pathway analysis.

Cell-based assay: Evaluates compound effects directly in living cells, providing physiological relevance for efficacy and toxicity testing.

Chemiluminescent assay: Utilizes chemiluminescent substrates to quantify biomolecular interactions or enzyme activities with high sensitivity.

ELISA assay: Detects and quantifies specific proteins or phosphorylated targets, supporting pathway and biomarker analysis.

Fluorescence resonance energy transfer (FRET) assay: Assesses molecular interactions or conformational changes by measuring energy transfer between fluorescent labels.

Fluorescent assay: Employs fluorescent tags or probes to detect cellular or biochemical activities, enabling multiplexed and sensitive detection.

Fluorescent polarization assay: Measures binding interactions based on changes in fluorescence polarization, useful for studying ligand-receptor or protein-protein interactions.

Homogeneous Time Resolved Fluorescence (HTRF) assay: Combines time-resolved fluorescence and FRET technologies for sensitive quantification of biomolecular events.

Luciferine/luciferase assay: Uses luciferase enzyme activity as a luminescent reporter for gene expression, cell viability, or signaling readouts.

Midazolam hydroxylation assay: Monitors cytochrome P450 activity via midazolam metabolism, relevant for drug metabolism and safety profiling.

Peptide as substrate: Employs synthetic peptides as enzyme substrates to measure kinase or protease activities.

Surface plasmon resonance assay: Provides label-free, real-time analysis of biomolecular binding kinetics and affinities, supporting target engagement studies.

We measure key pharmacological parameters that quantify the potency, binding affinity, and inhibitory strength of test compounds. These metrics, including IC-50, Kd, and Ki, are critical for comparing candidate efficacy and guiding lead optimization. Accurate parameter determination ensures informed decision-making in drug development pipelines.

IC-50: The concentration of a compound required to inhibit 50% of the target activity, serving as a standard measure of potency.

Kd: The dissociation constant representing the binding affinity between a drug and its target, with lower values indicating stronger binding.

Ki: The inhibition constant that quantifies the binding strength of an inhibitor to an enzyme, important for evaluating competitive inhibition and potency.

Recommended In Vitro Efficacy Tests

Akt Serine/Threonine Kinase 1

Akt Serine/Threonine Kinase 1 (AKT1) is a key downstream effector in the PI3K pathway, driving cell proliferation in Pik3Ca Related Overgrowth Spectrum (PROS). Accurate AKT1 testing is crucial for drug development targeting this pathway. Our service employs ATP and fluorescent assays, peptide substrates, BRET, and surface plasmon resonance to assess inhibitor potency and binding. Main parameters measured include IC-50 for inhibitory concentration and Kd for binding affinity.

Akt Serine/Threonine Kinase 2

Akt Serine/Threonine Kinase 2 plays a pivotal role in Pik3Ca Related Overgrowth Spectrum by mediating PI3K pathway signaling, driving abnormal cell growth. Testing Akt2 activity is crucial for drug development targeting this pathway. Our service utilizes peptide substrates, ATP assays, and fluorescent assays to measure kinase activity, with IC-50 determination as the primary parameter to assess compound efficacy and enable informed therapeutic development.

Akt Serine/Threonine Kinase 3

The Akt Serine/Threonine Kinase 3 testing service evaluates its key signaling role in Pik3Ca Related Overgrowth Spectrum, where dysregulation drives abnormal cell growth. This assay is essential for drug development targeting the PI3K/Akt pathway. Using peptide substrates, ATP consumption, and fluorescence-based detection, the assay quantifies kinase activity and determines drug efficacy, with IC-50 as the main parameter for inhibitor potency assessment.

Cytochrome P450 Family 3 Subfamily A Member 4

Cytochrome P450 Family 3 Subfamily A Member 4 (CYP3A4) metabolizes drugs used in Pik3Ca Related Overgrowth Spectrum (PROS) therapy, influencing efficacy and safety. Testing CYP3A4 activity is crucial to predict drug-drug interactions and optimize dosing. Our service utilizes the midazolam hydroxylation assay to assess CYP3A4 inhibition, providing key parameters such as IC-50 values for candidate compounds in PROS drug development.

Phosphatidylinositol-4,5-Bisphosphate 3-Kinase Catalytic Subunit Alpha

Phosphatidylinositol-4,5-Bisphosphate 3-Kinase Catalytic Subunit Alpha (PIK3CA) is a key driver of cellular overgrowth in Pik3Ca Related Overgrowth Spectrum (PROS) disorders. Accurate testing of PIK3CA activity is vital for drug development targeting PROS. Using assays such as luciferase, ATP, FRET, ELISA, chemiluminescence, cell-based, HTRF, and fluorescence polarization, we determine critical drug parameters including IC₅₀, Kᵢ, and K_d for potency and binding affinity assessment.

Phosphatidylinositol-4,5-Bisphosphate 3-Kinase Catalytic Subunit Beta

The Phosphatidylinositol-4,5-Bisphosphate 3-Kinase Catalytic Subunit Beta (PIK3CB) plays a critical role in Pik3Ca Related Overgrowth Spectrum by mediating PI3K signaling pathways involved in abnormal cell growth. Accurate PIK3CB testing is essential for drug development targeting these pathways. We offer high-throughput ATP, FRET, fluorescent, chemiluminescent, cell-based, HTRF, and fluorescence polarization assays, quantifying key parameters such as IC₅₀, Kᵢ, and Kd to assess inhibitor potency and binding affinity.

Phosphatidylinositol-4,5-Bisphosphate 3-Kinase Catalytic Subunit Delta

Phosphatidylinositol-4,5-Bisphosphate 3-Kinase Catalytic Subunit Delta is implicated in Pik3Ca Related Overgrowth Spectrum by dysregulating PI3K signaling, driving abnormal cell proliferation. Testing its activity is crucial for drug development targeting this pathway. We offer ATP, FRET, fluorescent, chemiluminescent, cell-based, HTRF, and fluorescent polarization assays, determining key parameters such as IC50, Ki, and Kd to evaluate compound potency and binding, supporting effective therapeutic discovery.

Phosphatidylinositol-4,5-Bisphosphate 3-Kinase Catalytic Subunit Gamma

Our Phosphatidylinositol-4,5-Bisphosphate 3-Kinase Catalytic Subunit Gamma (PIK3CG) testing service supports Pik3Ca Related Overgrowth Spectrum drug development by assessing PIK3CG's role in aberrant PI3K signaling implicated in overgrowth disorders. Utilizing ATP, FRET, chemiluminescent, HTRF, and fluorescent polarization assays, we quantify key inhibition and binding parameters—IC₅₀, Ki, and Kd—providing critical data for evaluating candidate drug efficacy and specificity targeting PI3K pathway dysregulation.