• in-vitro-tox

in vitro Biology

Pharmaron Biology provides a spectrum of high quality services to support various stages of drug discovery and development. With state-of-the art facilities and instrumentation, proprietary data management systems – CEDAR™, we offer services in assay development, compound screening, PD/biomarkers, bioreagents and structural biology, and in vitro toxicity.

We have a team of experienced scientists that are familiar with drug discovery processes, and understand how and where biology services can help our clients succeed in target validation, hit identification, hit to lead, lead optimization, and selection of preclinical candidates.  These services are provided as standalone services, or as a part of integrated projects that also include components such as medicinal chemistry, DMPK, in vivo pharmacology and toxicology. 

  • Experienced leadership team with drug discovery experience
  • Excellent team of cell biologists, molecular biologists, enzymologists, protein biochemists, structure biologists and electrophysiologists
  • State-of-the art facilities and instrumentations
  • Experience with major target classes
  • Rigorous assay development, validation, & execution
  • Track record of successful delivery of drug candidates for our partners

Our in vitro Biology team works closely with our partners in selecting and evaluating targets, assay format, and design flexible work flows to meet project specific goals. We have strong expertise in key therapeutic areas such as cell biology, oncology, immun-oncology, metabolic disorders, immunology/inflammation, neuroscience, histology/pathology.

Cell-Based Assays

Cell based assays are more close to in vivo conditions and are often more informative than biochemical assays on functional activities of test compounds. Pharmaron biology has over 400 human and mouse cell lines, and offers an extensive line of cell-based assays for target evaluation/identification, compound screen, biomarker analysis, and mode of action studies.  About 50% of assays developed, validated for compound screen are cell-based assays.

  • Sable cell line generation and characterization
  • Cell-based assay development
  • Primary and secondary cell-based functional assays for compound screening
  • Signal transduction assays (protein phosphorylation, nuclear translocation)
  • Target validation (siRNA, shRNA knockdown, Crispr-Cas9 gene editing)
  • Anti-proliferation/cytotoxicity cancer panel screening
  • Cell migration, apoptosis, cell cycle, angiogenesis assays
  • Standard cell-based assay formats with primary cells (such as PBMCs, primary hepatocytes, T cells, primary DRG neurons, etc.) and cell lines (HEK293, CHO, THP1, Ramos, Hep3B, etc.)
    • Growth inhibition/cytotoxicity assays
    • Transwell migration, chemotaxis assays
    • Reporter assays, cAMP, IP1, FLIPR
    • High-content screening using Acumen
    • Cytokine, chemokine secretion assay by ELISA, MSD multiplex,
    • Real-time qPCR and Western-blot, FACS
    • Transporter uptake assays

in vitro Biology for Oncology, immun-oncology

As cancers represent a large class of diverse diseases characterized by abnormal cell growth, ever increasing number of signaling molecules and pathways are being uncovered as regulators controlling cell growth, differentiation, and migration. We strive to stay current with the cutting edge of oncology research and continue to strengthen our comprehensive platform for cancer drug discovery. Our in vitro oncology team is experienced in target identification and validation. We have strong expertise in developing and optimizing diverse assays in targets ranging from kinases, GPCR, nuclear receptors, T-cell receptors, to metabolic enzymes, epigenetics enzymes, and oxygen sensors. We have provided compound profiling support to numerous medchem projects, also working closely with our in vivo oncology team to perform PKPD correlation studies, and providing data based input in model selection and study design for better translational success in efficacy studies.

  • Kinase panel screen (>300 kinases)
  • Epigenetic enzyme assays (HDAC, PARP, Histone methyl transferase assays)
  • T cell activation/proliferation assay
    • IDO, TDO assays
    • Mixed Lymphocyte Reaction (MLR)
    • PD1/PD-L1 binding assay
  • Cancer panel screen (>400 human/mouse cell lines, multiple readout)
  • Target specific mutant cell line construction
  • Cell proliferation assay (standard and long term) including combination studies
  • High content analysis for target protein phosphorylation, nuclear translocation
  • Clonogenic assay
  • Cell cycle analysis (cell cycle arrest, H3-thymidine incorporation assay)
  • Cell migration/invasion, apoptosis, angiogenesis assays
  • PK/PD biomarker analysis
    • Tissue, serum/plasma, CSF, excretion
    • RNA expression, Gene copy number, protein/peptide, protein modifications, small molecules
    • qPCR, Western Blot, ELISA/MSD, IHC, FACS, LC/MS
  • Hot spot cancer mutation analysis

in vitro Biology for Metabolic disorders

Metabolic diseases are modern society disorders caused by imbalance between energy intake and expenditure, manifesting in mis-regulated lipid and glucose metabolism. Despite numerous drugs are available to control diabetes, hypertension, and cardiovascular diseases, there are still great needs for better pharmacotherapies that improve efficacy and reduce side effects in the chronic treatment paradigm, and to meet unmet medical needs in the case of obesity and nonalcoholic steatoheptitis (NASH). Pharmaron offers a variety of in vitro biochemical and cell based assays targeting different classes of validated and novel metabolic drug targets. These in vitro assessments are supported by validated animal models in diabetes, obesity, hypertension, dyslipidemia, and thrombosis.

  • Glucose-stimulated Insulin Secretion (GSIS, rat islet, pancreatic cell lines)
  • Cell based Insulin signaling (Insulin stimulated pAKT assay)
  • GPCR assays (FLIPR Calcium flux, cAMP, β-arresting, GTPγs assays)
  • Nuclear hormone receptor assays (PPAR, FXR, RORgt, ER, PR, LXR, GR etc)
  • Receptor binding assays (filtration, TR-FRET, SPA)
  • Metabolic enzyme assays (ACC, ACL, mTOR, PTP1B, AMPK, PFKFB3, PDE etc)
  • Transporter assays (Glu4, Npt2B, URAT1, Cl/HCO3 Exchanger etc)
  • Cell based C14 fatty acid, cholesterol synthesis assays

Immunology/anti-inflammation Biology

Recently, significant progress has been made in understanding the link between immune-modulation and chronic inflammatory, auto-immune and oncological diseases.  For example, rheumatoid arthritis is an auto-immune disease characterized by chronic infiltration of B, T cells and macrophages into the joints, inflicting local cytokine reactions that lead to pain and destruction of joint cartilage and bones. The new insights have helped advance improved anti-inflammatory drugs by targeting newly identified mediators in the cytokine induction and signaling pathways. Pharmaron in vitro biology team has established many enzymatic and cell based assays, intervening innate and adaptive immune-response pathways.

    • Inflammation enzyme assays (JAK, BTK, P38, PIM, IRAK4, IDO  etc)
    • Whole blood cytokine production (human/rat)
    • LPS or other challenger-induced THP-1 cell cytokine production
    • Cell proliferation (PBMC/differentiated PBMC)
    • Cell based RORgt assay, TLR activation assays
    • CD marker evaluation by FACS
    • Cytokine/chemokine profiling (AlphaLISA, MSD, Luminex)

in vitro Biology for CNS Diseases

    • Patch clamp hERG activity
    • DRG neuron activation
    • Ion channel assays
      • Nav1.5
      • P2X3
      • P2X4
      • P2X7