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 ~400 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. In addition, biomarker discovery and development is integrated into every project, ensuring the co-development of pharmacodynamic, as well as patient stratification and toxicology biomarkers. Target validation, complex cellular screening, mechanism of action studies and early translational biology research complete the breadth of activities performed by the Evotec team in eukaryotic and prokaryotic systems. 

Evotec’s core expertise covers therapeutic areas such as CNS diseases, pain, inflammation & immunology, fibrosis, metabolic diseases, oncology and anti-infectives, and extends into a variety of rare diseases. The therapeutic 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. Building upon a modality-agnostic platform, the scientific team is also familiar with not only small molecule drug development, but also various biologic, gene therapy and cell therapy approaches.  With access to a world-class portfolio of assay technologies for biochemical and cellular assays, our drug discovery and disease biology expertise has been repeatedly proven to be a driver of success for our partner’s projects.

Read-outs and Assay technologies

To effectively answer scientific questions raised within projects, 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 read-outs to reveal the true mechanism of action of compounds to aid the selection, optimisation and/or validation of the best chemical, biological or cellular leads.

  • Classical receptor pharmacology and biochemistry: assess compound binding kinetics or reversibility via the application of radioactive binding assays, as well as enzymology expertise
  • Cell-based read-outs: recombinant systems, native cellular or tissue-based models are used for routine read-outs performed at Evotec, including immune assay formats (MSD, Singulex, FRET), flow cytometry and high-content imaging, in both 2D or 3D contexts
  • Electrophysiology: from high-throughput automated screening to rapid perfusion studies and slice electrophysiology, Evotec covers all the needs around ion channel drug discovery

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
  • 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
  • Genetic modification: unlocking gene (or drug) MoA is made possible with CRISPR (KO/KI), RNAi, overexpression, lentiviral and AAV tools applied to cells, tissues or in vivo

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

Biochemical and cell-based assay technologies

  • HTRF, FP, FRET, Alphascreen, FCS+plus
  • FLIPR direct flux and membrane potential dyes
  • Electrophysiology (Automated and Manual patch clamp, slice EPHYs) 
  • Interaction and reporter gene assays (nanoBret, InCell Pulse, nanoLuc, GFP etc) 
  • Immunoassays (MSD, SMCxPro, Luminex, Quanterix) 
  • Simple Western/Jess and In-Cell Western
  • Radiometric assays and autoradiography (3H, 125I, 59Fe, 32P, 35S)
  • Proliferation, apoptosis, autophagy and migration assays (Incucyte, transwell, etc) 
  • iPSC, primary cell, mixed culture and spheroid models
  • Whole blood assays 
  • Multiparametric flow cytometry and FACS
  • Cellular and ex vivo imaging (high-content assay, Incucyte, confocal, scanners) 
  • Metabolic analysis (Seahorse, metabolite analyser and metabolomics) 
  • CRISPR, RNAi, AAV and LV
  • High-throughput qPCR, RNAseq and scRNAseq
  • MS-based proteomics and metabolomics
  • Histology and immuno-histochemistry (including ISH/RNAscope/Basescope)
  • Access to biological samples of control and disease subjects

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, IN Cell Analyser) including a dedicated expert team focusing on the development of novel image analysis scripts.
High-content imaging-based assays (HCAs) enable the combination of complex cellular models with disease-relevant assay read-outs. This allows the modelling of disease in vitro, the bridging of the gap between recombinant cellular systems and in vivo studies, and even bridging into the clinic.

Synapse formation in rat primary neurons

HCA applications include the development and implementation of assays for phenotypic hit identification screens, but also disease-relevant cell-based assays for compound characterisation during H2L and lead optimisation. 
Evotec has successfully evaluated a variety of cellular model systems, including 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, including in the CNS area, using ex vivo histology and immuno-histochemistry 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, Qube 384 and Synchropatch 384PE for HTS through to manual rigs including specialisation for fast perfusion mechanistic studies. For translational analysis, Evotec offers several multi electrode array (MEA) platforms with primary and iPSC-derived neuronal cultures. Brain slice recordings are available from MEA, Slice Master & manual patch technologies. Epithelial tissues and small muscle tissue rings studies are accessible with Ussing chambers and Wire Myograph devices.

Throughput (compounds per week)
4,000 - 10,000
HTS, hit-to-lead, lead optimisation, early safety assessment
Qube 384
4,000 - 10,000
HTS, hit-to-lead, lead optimisation, early safety assessment
Q-patch HTX®
150 - 450
Hit-to-lead, lead optimisation, early safety assessment
Q-patch II
150 - 450
Hit-to-lead, lead optimisation,early safety assessment
Manual Electrophysiology (including fast perfusion)
1 - 25
Lead optimisation, safety pharmacology
Wire Myograph
8 parallel recordings
Lead optimisation
Ussing chamber
8 parallel recordings
Lead optimisation
Platform SynchroPatch384PE
Format 384-well
Throughput (compounds per week) 4,000 - 10,000
Applications HTS, hit-to-lead, lead optimisation, early safety assessment
Platform Qube 384
Format 384-well
Throughput (compounds per week) 4,000 - 10,000
Applications HTS, hit-to-lead, lead optimisation, early safety assessment
Platform Q-patch HTX®
Format 48-well
Throughput (compounds per week) 150 - 450
Applications Hit-to-lead, lead optimisation, early safety assessment
Platform Q-patch II
Format 48-well
Throughput (compounds per week) 150 - 450
Applications Hit-to-lead, lead optimisation,early safety assessment
Platform Manual Electrophysiology (including fast perfusion)
Throughput (compounds per week) 1 - 25
Applications Lead optimisation, safety pharmacology
Platform MEA
Format 96-well
Throughput (compounds per week) 200-500
Applications Hit-to-lead
Platform Wire Myograph
Format 8 parallel recordings
Throughput (compounds per week) varies
Applications Lead optimisation
Platform Ussing chamber
Format 8 parallel recordings
Throughput (compounds per week) varies
Applications Lead optimisation

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.

AMPA currents

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AMPA currents are recorded in rat cortical neurons in culture. The selective blocker NBQX inhibited the AMPA activated current.

AMPA currents are recorded in rat cortical neurons in culture. The selective blocker NBQX inhibited the AMPA activated current.

AMPA currents are recorded in rat cortical neurons in culture. The selective blocker NBQX inhibited the AMPA activated current.

AMPA currents are recorded in rat cortical neurons in culture. The selective blocker NBQX inhibited the AMPA activated current.

AMPA currents are recorded in rat cortical neurons in culture. The selective blocker NBQX inhibited the AMPA activated current.

AMPA currents are recorded in rat cortical neurons in culture. The selective blocker NBQX inhibited the AMPA activated current.

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) studies using a variety of leading technologies, including
    • Macromolecule synthesis assay (MMS) 
    • Whole-genome sequencing (WGS), RNASeq and TnSeq
    • Bacterial cytological profiling (BCP) and phenotypic microarrary (Biolog)
    • Mass spectrometry applied to metabolite or protein studies
  • Viruses: multiple assay formats to study viruses including plaque, CPE, neutralisation assays

If you are interested in

  • Our capabilities in anti-infectives, please click here
  • Our in vivo pharmacology capabilities for anti-infectives, please clicke here

In vitro Biology for autoimmune and inflammation 

Evotec has a strong expertise in supporting autoimmune and inflammation and immunology drug discovery projects, starting from target validation and 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 T-cell subsets, B-cells, neutrophils and monocyte lineages
  • Expertise in a variety of target classes including ion channels, kinases, transporters, cytokine PPI, GPCRs, enzymes and pattern recognition receptors. Example target classes include JAKs, purinoceptors, CRTH2, TNF/TLR families and NF-kB signalling
  • Broad technology platform to support biochemical assays, functional cell based assays and phenotypic characterisation of drug responses, including high-content read-outs, FACS cytometry with 384 well sampler and MSD or Luminex
  • Development and validation of novel customised assays for screening, validation and drug development. Some examples include:
    • T-cell expansion
    • Cytokine profiling
    • Cell surface receptor expression
    • Neutrophil extracellular traps (NET) formation
    • T-, B- ,macrophage and dendritic cell activation
    • In vitro fibroblast to myofibroblast transition

If you are interested in

  • Our capabilities in inflammation and immunology, please click here
  • Our in vivo pharmacology capabilities for inflammation and immunology, please clicke here

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

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

If you are interested in

  • Our capabilities in metabolic diseases and complications, please click here
  • Our in vivo pharmacology capabilities for metabolic diseases and complications, please clicke here

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

If you are interested in

  • Our capabilities in neurosciences, please click here
  • Our in vivo pharmacology capabilities for neurosciences, please clicke here

In vitro Biology for Oncology

Evotec has significant expertise in a broad range of oncology areas including signal transduction, cancer metabolism, tumour microenvironment, immuno-oncology, tumour metastasis and vascularisation, DNA damage and epigenetics. As a result, the Evotec team integrates assays based on cancer and/or microenvironment related cells (fibroblasts, immune cells, endothelial cells, or adipocytes) in 2D, co-culture or 3D culture to recapitulate tumour and microenvironment conditions.  In addition, specific assays to study targeted therapy resistance, drug impact on cancer metabolism, or cancer-specific phenotypic responses have been applied in project-specific contexts. Examples include:

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)

  • Characterisation of tumour microenvironment (scRNAseq, multi-colour IHC, flow cytometry)
  • Metabolism (SeaHorse, Oxygraphy, Hypoxia, Glycolytic/OXPHOS ATP, metabolite consumption/ production)
  • DNA damage repair studies, including synthetic lethality, strand displacement, primer extension, DBR, HCA and end joining reporter assays
  • Flow Cytometry to support immuno-oncology (18 colour, cell to platelet analysis, FACS, ELISpot)
  • Organotypic approaches for biomarker and MoA validation  on fresh patient samples
  • Target identification approaches e.g. synthetic lethality via phenotypic, RNAi or CRISPR screens

If you are interested in

  • Our capabilities in oncology, please click here
  • Our in vivo pharmacology capabilities for oncology, please clicke here

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