In vitro biology


The cornerstone of Evotec’s in vitro biology function is disease and target biology expertise coupled with state-of-the-art technology platforms. A large team of >100 scientists with extensive industrial experience supports the in vitro pharmacological characterisation of compounds as part of hit expansion, lead finding and lead optimisation projects, generating project-relevant high-quality data in short turnaround times. The team also routinely support in vivo studies with pharmacodynamic read-outs and engage in early translational biology research. In addition to its extensive capabilities in mammalian biological systems, Evotec has in-depth in vitro microbiology expertise spanning a broad range of pathogens from bacteria to fungi and viruses.

Evotec’s core expertise covers areas such as CNS diseases, pain, inflammation & immunology, metabolic disease, oncology and anti-infectives. Evotec’s disease area expertise is combined with in-depth know-how in relevant target classes, including classical target families such as GPCRs, ion channels and kinases, as well as a large diversity of other target areas such as transporters, protein-protein interactions and multiple enzyme families. Beyond biology and drug discovery know-how, Evotec has access to a world-class portfolio of assay technologies for biochemical and cellular assays that have shown to be drivers for the success of our partner’s projects.

Our scientific expertise and understanding of disease mechanisms combined with our track record in setting up relevant in vitro models and translational biomarker assays are a key factor in our success when working with our partners.

Read-outs and Assay technologies

For each target, approach and philosophy, Evotec utilises the most appropriate technology coupled to disease-relevant assay design. With a plethora of biochemical, biophysical and cellular assay technologies, Evotec scientists will combine many readouts to reveal the true mechanism of action of compounds to aid the selection and optimisation of the best chemical starting points.

  • Classical receptor pharmacology: assess compound binding kinetics or reversibility via the application of radioactive binding assays
  • Cell-based technologies: using recombinant systems and native cellular or tissue-based models, routine read-outs performed at Evotec include immune assay formats (MSD, Singulex, FRET) through to flow cytometry and high-content imaging

Monitoring fibrosis through high-content imaging in lung cells to support compound optimisation

  • Label-free approaches: define and optimise the mode of binding via a combination of LC/MS NMR, ITC, DSF, SPR & MST methods
  • Electrophysiology: from high-throughput automated screening to rapid perfusion studies and slice electrophysiology, Evotec covers all the needs around ion channel drug discovery
  • Phenotypic screening: Evotec´s origins in the development of HCS imaging hardware persists with a dedicated imaging team, providing and developing image analysis tools supporting Evotec´s phenotypical imaging platform

As an integral part of Evotec´s drug discovery platform each platform is served via a professional compound handling and data management infrastructure.

Biochemical and cell-based assay technologies

  • HTRF, FP, FRET, Alphascreen
  • FCS+plus
  • Membrane potential dyes
  • Electrophysiology (High-troughput, manual, slice EPHYs)
  • Reporter gene assays (Luciferase, GFP variants)
  • Immunoassays (TR-FRET, MSD, Singulex)
  • Radioactive assays (3H, 125I, 59Fe, 32P)
  • Migration assays (Incucyte)
  • Primary cell-based models
  • Whole blood assays
  • Spheroid models
  • iPS models
  • Flow cytometry
  • Cellular and ex vivo imaging (high-content assays)
  • Metabolic analysis (Seahorse, metabolites analyser and metabolomics)
  • High-throughput qPCR

Biophysical assay technologies

  • Surface plasmon resonance (SPR)
  • Mass spectrometry (LC-MS)
  • Thermal shift (differential scanning fluorimetry)
  • Microscale thermophoresis (MST)
  • Isothermal titration calorimetry (ITC)

Phenotypic assays

Evotec is a world leader in performing cellular and tissue-based imaging assays. An extensive infrastructure is available with state-of the art imaging devices (Phenix, Opera, Operetta, ArrayScan) including a dedicated expert team focusing on the development of novel image analysis scripts.
Imaging-based assays are enabling as they allow to combine complex cellular models with disease-relevant assay read-outs. This allows to model disease in vitro and to bridge the gap between recombinant cellular systems and in vivo studies but also to the clinic.

Synase formation in rat primary neurons

Applications include the development and implementation of assays for phenotypic hit identification screens, but in particular to apply disease-relevant cell-based assays for compound characterisation during H2L and lead optimisation.
Cellular models include primary neuronal cultures, kidney cells, immune cells, tumour cells, lung cells, muscle & beta cells as well as patient derived stem cells. Furthermore, imaging is a key platform for tissue-based target validation activities in the CNS area, using ex vivo histology and immunohistochemistry approaches.

Ion Channels

Evotec´s dedicated ion channel  facility represents a synergy of experienced electrophysiologists with drug discovery expertise  and state-of-the-art hardware for both HTS as well as biophysical analysis of channel activity brought together to address the challenge of ion channel drug discovery.

Hardware platform

Regardless of the ion channel of interest, ligand- or voltage-gated, Evotec offers the appropriate platform including FLIPR and Synchropatch 384PE for HTS through to Dynaflow equipped manual rigs for fast perfusion mechanistic studies. For translational analysis, Evotec now offers the multi electrode array ("MEA") platform with primary and iPSC-derived neuronal cultures.

Compound profiling

Numerous medicinal chemistry programmes are supported by the ion channel group providing weekly turnaround times and project leadership. Using Evotec´s compound management facility linked to informatics tools for data management and upload to client´s databases, a professional process is offered from start to finish.

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Voltage-gated sodium channels (NaV) are expressed in neurons. The selective NaV blocker, TTX (300nM) inhibited NaV signal in big diameter cells.

Voltage-gated sodium channels (NaV) are expressed in neurons. The selective NaV blocker, TTX (300nM) inhibited NaV signal in big diameter cells.

Tissue profiling and primary cell analysis

As programmes progress through the drug discovery process, a clear understanding of target engagement in disease-relevant tissue is critical for predicting both in vivo efficacy as well as future human dose. With in-house preparation of primary tissue such as DRGs, NDGs, human iPSC-derived cell types including tissue slices, the effects of compounds on neural networks is assessed.

In vitro Biology for Anti-infectives

MICROBIOLOGY – Bacteriology, Mycology, Virology, Parasitology

Evotec’s infectious disease group boasts state-of-the-art microbiology facilities and has a full range of in vitro assay capabilities in order to study the following pathogens:

  • Bacteria including Gram positives and Gram negatives covering ESKAPE pathogens
  • Fungi including Candida, Aspergilli, and others
  • Viruses including Respiratory Syncytial Virus (RSV)
  • Parasites including Toxoplasmosis gondii

Evotec has a comprehensive strain bank, EVOStrAIn™, which is a constantly evolving resource of strains and clinical isolates of bacteria, fungi, viruses and parasites, many of which are validated in in vivo models of infection. Isolates are highly characterised and, in many cases, mechanisms of resistance defined. EVOStrAIn™ contains an extensive range of geographically diverse human bacterial and fungal pathogens that cover isolates susceptible and resistant to current antimicrobial drugs.

Core strengths of Evotec’s microbiology capability:

  • Full HTS: 384- and 1536-well format. BSL-2 containment for phenotypic and growth inhibition assays. Target based screening and multiple read-outs including fluorescence, luminescence, optical density, and HCS
  • In vitro microbiology: detailed characterisation of antimicrobials including MIC and MBC/MFC determination, intracellular killing, frequency of resistance, timekill and PAE studies using single or combinations of agents. Industry-standard methods such as CLSI, EUCAST and BSAC.  Bespoke methods developed where required
  • Hollow fibre PK/PD or bioreactor human cell systems for detailed profiling and characterisation of novel anti-infective agents, and compound/drug combination studies for assessment of synergistic, antagonistic and additive effects
  • Biofilms: characterisation of compounds and their ability to disrupt biofilms. Multiple assay formats
  • Mechanism of action (MOA) determination. Resistant mutant generation and screening of mutant libraries. State-of-the-art proteomics and biomarker platform for cellular target profiling
  • Viruses: multiple assay formats to study viruses including plaque, CPE, neutralisation assays

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Our capabilities in anti-infectives In vivo pharmacology for anti-infectives

In vitro Biology for inflammation and Immunology

Evotec has a strong expertise in supporting inflammation and immunology drug discovery projects from target validation, in vitro proof of concept through to pre-clinical candidate nomination:

  • Regular profiling in biochemical and functional cellular assays including primary human lymphocytes and whole blood assays, including:
    • Human primary PBMCs
    • CD4+, CD8+ T-cells, and subsets
    • B-cells
    • Neutrophils
    • Monocyte and macrophages
  • Portfolio of relevant in vitro models for multiple target classes including ion channels, kinases, transporters, cytokine PPI, GPCRs, enzymes and pattern recognition receptors. Example target classes include:
    • MAP kinase/ERK family
    • JAKs
    • Purinoceptors
    • CRTH2
    • Tryptophan metabolism, Prostaglandin synthesis pathways, other enzymes
    • TNF family
    • Pattern recognition receptors and pathways
  • Broad technology platform to support biochemical assays, functional cell based assays and phenotypic characterisation of drug responses
    • High-content read-outs
    • FACS cytometry w/384 well sampler
    • TR-FRET
    • MSD
    • Broad in vitro assay platform from enzymatic assays to FLIPR
    • Receptor ligand binding studies
    • Biophysical assay platform including SPR, NMR, LC-MS, DSF
  • Development and validation of novel customised assays for screening, validation and drug development. Example assay formats:
    • T-cell expansion
    • Cytokine profiling
    • Cell surface receptor expression
    • NET formation
    • T-, B- and macrophage activation
    • In vitro fibroblast to myofibroblast transition

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Our capabilities in immunology and inflammation In vivo pharmacology for immunology and inflammation

In vitro Biology for Metabolic Diseases and complications

Evotec has significant experience in a broad range of metabolic disorders including diabetes and diabetic complications such as kidney disease and NASH. Beyond our expertise in classical drug targets such as GPCRs and enzymes, a significant effort has been put into building disease-relevant cellular assays to support target discovery, target validation and for compound characterisation.

Examples include:

  • Skeletal muscle regeneration assays using cells from muscular dystrophy patients or animal models
  • Assays monitoring protection of human podocytes or freshly isolated pig/rat glomeruli
  • Assays measuring anti-fibrotic effects across different primary cell types
  • Assays studying GI biology

Myotube formation from human muscle cells

  • Comprehensive set of primary islet cell assays covering all relevant aspects of pancreatic beta cell turnover and function
    • Protection/reversal of metabolic stress-induced beta cell de-differentiation
    • Cell specific primary rat and human beta cell replication (plated or intact islets)
    • Cell specific quantification of beta cell apoptosis in plated islets
    • Insulin secretion with rat and human islets

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Our capabilities in metabolic diseases and complications In vivo pharmacology for metabolic diseases and complications

In vitro Biology for Neurosciences

Evotec has built significant expertise in a range of neuroscience research areas over the last 15 years including neurodegeneration, psychiatric diseases, neuroinflammation and pain. Currently over 100 FTEs are dedicated to research in this area. 

Main capabilities and activities include:

  • Extensive pharmacology expertise across relevant target classes: GPCRs, ion channels, enzymes and others investigating potency, selectivity, mechanism of action and receptor occupancy
  • World-class platform for ion channel drug discovery, ranging from slice electrophysiology and manual patch clamp studies to fully automated screening

Outgrowth of dorsal root ganglia

  • Portfolio of disease-relevant secondary assays using primary neurons, microglia, astrocytes and co-cultures, stem cells and slice cultures
  • State-of-the-art imaging platform analysing synapse and spine morphology
  • Whole tissue radio-labelled ligand binding and competition studies
  • World-class target validation platform encompassing in vivo target modulation followed by systematic, imaging-based ex vivo analysis
  • Target engagement and biomarker assays with a focus on Huntington’s disease

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Our capabilities in neurosciences In vivo pharmacology for neurosciences

In vitro Biology for Oncology

Evotec has a significant expertise in a broad range of oncology areas including signal transduction, cancer metabolism, tumour microenvironment, immuno-oncology, tumour metastasis and vascularisation, apoptosis and epigenetics. All our projects benefit from an outstanding know-how in specialised drug discovery and an extensive portfolio of molecular biology, biochemical, biophysical and cellular assay systems to progress.

Using state-of-the-art technologies, Evotec developed integrated assays based on cancer cells or microenvironment related cells (fibroblasts, immune cells, endothelial cells, or adipocytes) in 2D, co-culture or 3D culture to recapitulate tumour and microenvironment conditions and specific assays to study targeted therapies resistance, drug impact on cancer metabolism, or microenvironment (immune cells, fibroblasts, adipocytes, endothelial cells):

A549 lung cancer cells

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A549 lung cancer cells labelled with Hoechst (blue), mitotracker (orange), phalloidin (green) and lysotracker (deep red)

A549 lung cancer cells labelled with Hoechst (blue), mitotracker (orange), phalloidin (green) and lysotracker (deep red)

  • Metabolism (SeaHorse, Oxphography, Hypoxia, Glycolytic/ OXPHOS ATP, metabolites consumption/ production)
  • Signalling
  • Proliferation, apoptosis, migration, invasion models
  • Immunohistochemistry
  • Flux Cytometry to support immuno-oncology (9 colors cytometry, Cell to platelet analysis, Cell sorter)
  • Organotypic approach: connection with translational group and clinics in order to validate biomarkers developed in vitro on human fresh samples
  • mAb evaluation know-how in vitro strongly connected with biophysical methods
    • Antibody characterisation (Affinity constants determination, selectivity, cross-reactivity, Epitope mapping)
    • Antibody efficacy (PPI, signalling, cross reactivity, cellular functional assays)
  • Target identification approaches via phenotypic, RNAi or CRISPR screens
  • Customised high-content imaging assays for phenotypic screening through to supporting lead optimisation

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Our capabilities in oncology In vivo pharmacology for oncology

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