In Vitro Efficacy Testing Services for Metastatic Colorectal Cancer

We provide robust and sensitive in vitro screening and characterization platforms for accelerating the discovery and screening of potential therapies for Metastatic Colorectal Cancer. Our service offers comprehensive profiling of candidate drugs, biologics, and immunotherapies to elucidate their efficacy, mechanism of action, and selectivity in relevant cellular and molecular systems. Key targets and pathways evaluated include EGFR, VEGF, RAS, BRAF, PD-1/PD-L1, and Wnt/β-catenin signaling, which are central to colorectal cancer progression and metastasis. Our assays are designed to assess cellular proliferation, apoptosis, immune checkpoint interactions, and other pathological processes critical to tumor growth and therapy resistance.

We offer a diverse suite of in vitro testing methods, including biochemical assays, cell-based functional assays, binding assays, and advanced detection technologies. These platforms enable quantitative and qualitative evaluation of drug efficacy, target engagement, and pathway modulation in relevant colorectal cancer models. Our methods support comprehensive characterization from early screening to detailed mechanistic studies.

ATP assay: Measures cellular ATP levels to assess cell viability, proliferation, and cytotoxicity in response to candidate compounds.

ATP assay (at 0.01 mM): Evaluates the impact of low-concentration compounds on cell metabolism and viability.

ATP assay (at 1 mM): Assesses compound effects at higher concentrations to determine dose-dependent responses.

Biolayer interferometry assay: Real-time, label-free analysis of biomolecular interactions, such as drug-target binding kinetics.

CHO-K1 Chinese hamster ovary cells transfected with PDL1/OKT3: Functional cell-based assay to evaluate immune checkpoint inhibitor activity via engineered cell lines.

Cells transfected with PD1/NFAT/luciferase: Reporter assay to monitor PD-1/PD-L1 pathway inhibition and downstream signaling.

Chemiluminescent assay: Sensitive detection of enzymatic activity or biomarker expression using light emission.

Competitive binding assay (qPCR): Quantifies drug-target binding by measuring displacement of labeled ligands and downstream gene expression.

Competitive binding assay (with CD274): Assesses binding affinity and specificity of compounds targeting PD-L1 (CD274).

ELISA assay: Quantitative measurement of proteins, cytokines, or antibodies in cell supernatants or lysates.

Flow cytometry assay: Multiparametric analysis of cell surface markers, apoptosis, and intracellular proteins in heterogeneous populations.

Fluorescent assay: Detects cellular or molecular events using fluorescence readouts for high sensitivity.

Fluorescent polarization assay: Measures molecular interactions and binding affinities based on changes in fluorescence polarization.

Fluorescent-activated cell sorting (FACS) assay: Isolates and analyzes specific cell populations based on fluorescent labeling.

Homogeneous Time Resolved Fluorescence (HTRF) assay: Non-radioactive, high-throughput assay to detect protein-protein interactions and signaling events.

Jurkat human T-cell leukemia cells transfected with PD1/NFAT/luciferase: Functional immune assay for evaluating T-cell activation and immune checkpoint modulation.

Luciferine/luciferase assay: Measures gene expression or cell viability through bioluminescence emitted by luciferase enzyme activity.

Poly(glutamine/tyrosine) peptide as substrate: Utilized in enzymatic assays to study protease activity and substrate specificity.

RNA assay: Quantifies gene expression changes in response to drug treatment, providing insights into molecular mechanisms.

Radioactivity assay: Detects radiolabeled molecules to study binding, uptake, or metabolic activity with high sensitivity.

Surface plasmon resonance assay: Real-time analysis of molecular binding kinetics and affinity without the need for labeling.

We measure a comprehensive set of pharmacological parameters, including potency, efficacy, binding affinity, and minimal effective or inhibitory concentrations. These parameters are essential for comparing candidate therapies, optimizing dosing strategies, and understanding mechanisms of action. Accurate quantification of these metrics supports decision-making in drug development and prioritization.

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

IC-50: The concentration at which a compound inhibits a specific biological process or target by 50%, used to assess inhibitory activity.

Kd: The equilibrium dissociation constant, reflecting the binding affinity between a drug and its target; lower Kd values indicate stronger binding.

MEC: Minimal Effective Concentration, the lowest concentration at which a compound elicits a detectable biological effect, critical for dosing considerations.

MIC: Minimal Inhibitory Concentration, the lowest concentration of a compound that prevents detectable growth or activity, especially relevant for cytostatic agents.

pIC-50: The negative logarithm of the IC-50 value, allowing for easier comparison of compound potency on a logarithmic scale.

Recommended In Vitro Efficacy Tests

B-Raf Proto-Oncogene, Serine/Threonine Kinase

B-Raf Proto-Oncogene, Serine/Threonine Kinase plays a crucial role in metastatic colorectal cancer by driving abnormal cell signaling and tumor growth. Testing for B-Raf activity is vital for targeted drug development and personalized therapies. Our service employs ELISA and chemiluminescent assays to accurately assess B-Raf inhibition, measuring key parameters such as Minimum Inhibitory Concentration (MIC) and half-maximal inhibitory concentration (IC-50) to evaluate compound efficacy.

Cytotoxic T-Lymphocyte Associated Protein 4

Cytotoxic T-Lymphocyte Associated Protein 4 (CTLA-4) regulates immune responses and is implicated in immune evasion in metastatic colorectal cancer. Testing CTLA-4 is crucial for developing targeted immunotherapies. Our service utilizes flow cytometry, ELISA, and surface plasmon resonance assays to assess CTLA-4 expression, binding affinity (Kd), and inhibitory concentration (IC-50), providing essential data for drug efficacy and candidate selection in colorectal cancer drug development.

Dna Topoisomerase I

DNA Topoisomerase I plays a crucial role in DNA replication and repair, making it a key target in metastatic colorectal cancer drug development. Testing its activity is vital for evaluating drug efficacy and resistance. Our service utilizes a sensitive chemiluminescent assay to quantitatively assess enzyme inhibition. The main parameter measured is the Minimum Inhibitory Concentration (MIC), enabling precise determination of candidate drug potency against Topoisomerase I.

Epidermal Growth Factor Receptor

Epidermal Growth Factor Receptor (EGFR) plays a crucial role in driving tumor growth and progression in metastatic colorectal cancer (mCRC). EGFR testing is essential for identifying patients likely to benefit from targeted therapies. Our service utilizes advanced methods—including FACS, flow cytometry, ELISA, surface plasmon resonance, and ATP assays—to assess EGFR activity and inhibitor efficacy. Key parameters measured include IC-50, MIC, Kd, pIC-50, and MEC for robust drug development insights.

Fms Related Receptor Tyrosine Kinase 1

Fms Related Receptor Tyrosine Kinase 1 (FLT1/VEGFR1) promotes angiogenesis and tumor progression in metastatic colorectal cancer. Testing FLT1 activity is crucial for drug development targeting tumor growth and metastasis. Our service utilizes HTRF, ELISA, ATP, and radioactivity assays to accurately measure FLT1 inhibition. The primary parameter reported is IC-50, providing essential data for evaluating candidate drug potency and guiding therapeutic advancement.

Fms Related Receptor Tyrosine Kinase 4

Fms Related Receptor Tyrosine Kinase 4 (FLT4/VEGFR-3) plays a crucial role in lymphangiogenesis and metastasis in metastatic colorectal cancer. FLT4 testing is essential for identifying patients who may benefit from targeted therapies and for monitoring disease progression. Key methods include immunohistochemistry and quantitative PCR. Main parameters assessed are FLT4 expression levels and gene amplification status in tumor tissue samples.

Kinase Insert Domain Receptor

The Kinase Insert Domain Receptor (KDR/VEGFR-2) is a key driver of angiogenesis in metastatic colorectal cancer, promoting tumor growth and spread. Testing KDR activity is crucial for developing targeted therapies. Our service utilizes advanced methods—including chemiluminescent, HTRF, ATP, radioactivity, luciferase, competitive binding (qPCR), RNA, and ELISA assays—using poly(glutamine/tyrosine) substrates. Key parameters measured include pIC-50, MIC, and IC-50, providing comprehensive insights for drug development.

Kit Proto-Oncogene, Receptor Tyrosine Kinase

The Kit Proto-Oncogene, Receptor Tyrosine Kinase (KIT) plays a role in tumor progression and therapeutic response in metastatic colorectal cancer. KIT testing is crucial for identifying patients who may benefit from targeted therapies and for optimizing drug development. Key methods include immunohistochemistry (IHC) and next-generation sequencing (NGS). Main parameters assessed are KIT expression levels, gene mutations, and amplification status.

Programmed Cell Death 1

The Programmed Cell Death 1 (PD-1) pathway is crucial in immune evasion by metastatic colorectal cancer, making it a key target in drug development. PD-1 testing evaluates drug efficacy by measuring binding and inhibition using methods such as FACS, flow cytometry, ELISA, chemiluminescent and fluorescent assays, competitive binding, and surface plasmon resonance. Key parameters assessed include EC-50, IC-50, Kd, and MIC, providing vital data for immunotherapy candidate selection.