In Vitro Biology

The cornerstone of our in vitro biology function is disease and target biology expertise coupled with state-of-the-art technology platforms. A large team of >500 scientists with extensive industrial experience support the in vitro pharmacological characterization of compounds as part of hit expansion, lead finding and lead optimization projects, generating project-relevant high-quality data in short turnaround times. In addition, biomarker discovery and development are 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 our team in eukaryotic and prokaryotic systems.

Our 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, oligonucleotide, 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 & Assay Technologies

To effectively answer scientific questions raised within projects, we utilize the most appropriate technology coupled to disease-relevant assay design. With a plethora of biochemical, biophysical and cellular assay technologies, our scientists will combine many read-outs to reveal the true mechanism of action of compounds to aid the selection, optimization 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, we cover all needs around ion channel drug discovery 
  • Label-free approaches: define and optimize the mode of binding via a combination of LC/MS NMR, ITC, DSF, SPR & MST methods 
  • Phenotypic screening: Our origins in the development of HCS imaging hardware persists with a dedicated imaging team, providing and developing image analysis tools supporting our 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 our drug discovery offering, each platform is served by a professional compound handling and data management infrastructure. 

Biochemical & 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, HiBit 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) 
In Vitro Biology Read Outs And Assay Technologies

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

Phenotypic Assays

We are 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, Incucyte) 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. 

HCA applications include the development and implementation of assays for phenotypic hit identification screens, but also disease-relevant cell-based assays for compound characterization during H2L and lead optimization. 

We have successfully evaluated a variety of cellular model systems, including primary neuronal cultures, kidney cells, immune cells, tumor 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. 

In Vitro Biology Phenotypic Assays

Synapse formation in rat primary neurons


Our dedicated electrophysiology 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 activity brought together to address the challenge of ion channel and transporter drug discovery.

Hardware Platform

Regardless of the ion channel of interest, ligand- or voltage-gated, we offer the appropriate platform including FLIPR, Qube 384 and Synchropatch 384PE for HTS through to manual rigs including specialization for fast perfusion mechanistic studies. For translational analysis, we offer 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 using chambers and Wire Myograph devices.

Compound Profiling

Numerous medicinal chemistry programs are supported by the ion channel group providing weekly turnaround times and project leadership. Using our 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. 

Tissue Profiling & Primary Cell Analysis

As programs 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

Our 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

We have 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 characterized 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 capabilities:

  • 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 characterization of antimicrobials including MIC and MBC/MFC determination, intracellular killing, frequency of resistance, time kill and PAE studies using single or combinations of agents. Industry-standard methods such as CLSI, EUCAST and BSAC.  Bespoke methods developed where required 
  • Hollow fiber PK/PD or bioreactor human cell systems for detailed profiling and characterization of novel anti-infective agents, and compound/drug combination studies for assessment of synergistic, antagonistic and additive effects 
  • Biofilms: characterization of compounds and their ability to disrupt biofilms. Multiple assay formats 
  • Mechanism of action (MOA) studies using a variety of leading technologies, including CRISPR
  • Macromolecule synthesis assay (MMS)  
  • Whole-genome sequencing (WGS), RNASeq and TnSeq 
  • Bacterial cytological profiling (BCP) and phenotypic microarray (Biolog) 
  • Mass spectrometry applied to metabolite or protein studies 
  • Viruses: multiple assay formats to study viruses including plaque, CPE, neutralization assays 

In Vitro Biology For Autoimmune & Inflammation

We have 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 signaling 
  • Broad technology platform to support biochemical assays, functional cell-based assays and phenotypic characterization of drug responses, including high-content read-outs, FACS cytometry with 384 well sampler and MSD or Luminex 
  • Development and validation of novel customized 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 

In Vitro Biology For Metabolic Diseases & Complications

We have a 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 characterization.

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 
  • 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 
In Vitro Biology Metabolic Diseases Complications

Myotube formation from human muscle cells

In Vitro Biology for Neurosciences

We have 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 
  • 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 analyzing 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 
In Vitro Biology Neurosciences

Outgrowth of dorsal root ganglia

In Vitro Biology for Oncology

We have significant expertise in a broad range of oncology areas including signal transduction, cancer metabolism, tumor microenvironment, immuno-oncology, tumor metastasis and vascularization, 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 tumor 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: 

  • Characterization of tumor microenvironment (scRNAseq, multi-color 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 (28 colors, 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 
In Vitro Biology Oncology

A549 Lung cancer cells