SLAS Europe 2025

Iterative Screening – Unlocking High Throughput Screening Opportunities for Challenging Assays

^{Date and Time: Wednesday, May 21, 2025: 4:30 PM–5:30 PM CEST (Presentation Number: 1040-A)
Location: CCH – Congress Center Hamburg – Exhibition Hall
Presented by: Benedikt Bauer, Senior Data Scientist}

Iterative screening provides an alternative to full library screening. It uses machine learning to select smaller batches of compounds for physical screening, the data from which can refine the model and select the next subset for screening. It offers advantages over classical High-Throughput Screening (HTS) by achieving higher hit rates with fewer compounds and allows adjustments based on intermediate results. This method is particularly valuable for complex assays with limited materials, such as iPSC screens. The machine learning model applied in this study included compound structures, public domain HTS fingerprints, and in-house cell painting fingerprints. Retrospective analysis showed that iterative screening achieved over three times as many hits with similar numbers of screened compounds. In summary, iterative screening can provide an efficient, cost-effective and adaptable approach to drug discovery and is especially effective in the case of complex phenotypic readouts where traditional HTS may be prohibitively expensive.

A Cell Painting Based Assay for In Vitro Safety/Tox Screening

^{Date and Time: Wednesday, May 21, 2025: 4:30 PM–5:30 PM CEST (Presentation Number: 1084-A)
Location: CCH – Congress Center Hamburg – Exhibition Hall
Presented by: Daja Ruhlandt, Research Scientist Imaging and Analysis}

Evotec has developed a Cell Painting assay to assess compound safety early in the discovery process. This imaging-based phenotypic method uses fluorescent dyes to stain cellular structures and extracts numeric features from images to identify bioactivity. The assay, inspired by the US EPA's 2020 work, helps infer points of departure (PoDs) for subtle phenotypic changes to cell death. Continuous improvements are applied with new metrics for PoD definitions and machine learning for better classification. Future work will refine workflows and integrate ML-assisted processes into routine applications.

CRISPR Screening Platform for Identification and Validation of Novel Immune Cell Modulating Targets for I&I and I/O

^{Date and Time: Wednesday, May 21, 2025: 4:30 PM–5:30 PM CEST (Presentation Number: 1110-A)
Location: CCH – Congress Center Hamburg – Exhibition Hall H
Presented by: Bianca Giuliani, Research Associate, In Vitro Biology}

Evotec has developed a CRISPR screening platform using primary human CD4+ T cells to identify and validate new immune-modulating targets for autoimmune/inflammatory (I&I) and immuno-oncology (I/O) therapies. A druggable genome-scale knockout screen of over 8,000 genes revealed novel regulators of Tregs, with gene KOs that either increased (I&I) or decreased (I/O) FoxP3+ Tregs. Around 450 I&I hits were prioritized using Evotec’s proprietary Molecular Patient Database (E.MPD) and 54 targets were selected for downstream CRISPR-based target validation screening using orthogonal disease-relevant assays. NGS-based rhAmpSeq technology was used to correlate phenotype with genotype. Confirmed targets present starting points for new Treg-targeted therapies for autoimmune diseases and cancer.

Developing Life-Saving Oligonucleotide-Based Therapies for H-ABC Leukodystrophy

^{Date and Time: Wednesday, May 21, 2025: 4:30 PM–5:30 PM CEST (Presentation Number: 1114-A)
Location: CCH – Congress Center Hamburg – Exhibition Hall H
Presented by: Yalda Sedaghat, Senior Research Scientist, In Vitro Biology}

Evotec, in collaboration with SynaptixBio, are developing antisense oligonucleotides (ASOs) to treat H-ABC Leukodystrophy, a condition causing progressive motor skill decline in children. These ASOs target TUBB4A mRNA to repress gene expression. Due to the blood-brain barrier, ASOs are injected intrathecally into the cerebrospinal fluid. Evotec uses a combination of assays to screen for potency and safety, predicting in vivo efficacy and toxicity. For instance, assessing acute neurotoxicity through the use of primary neuronal cultures in combination with microelectrode array (MEA) technology offers a robust in vitro alternative that can significantly contribute to the reduction of animal use in neurotoxicological research.

High-Throughput Mass Spectrometry Workflows to Identify Covalent Binders: A Cysteine-containing Enzyme Case Study

^{Date and Time: Wednesday, May 21, 2025: 4:30 PM–5:30 PM CEST (Presentation Number: 1140-A)
Location: CCH – Congress Center Hamburg – Exhibition Hall
Presented by: Julien Parra, Senior Research Scientist} 

Covalent drugs have gained attention for their ability to target previously "undruggable" proteins. These drugs offer advantages like prolonged action, enhanced potency, and lower dosing requirements. Traditionally, concerns have been raised over potential off-target effects and toxicity, however, with advances in technologies and drug design, it has been possible to improve the selectivity profile of covalent drugs and to reduce the risk of adverse effects. In this respect, the use of covalent approaches has been valuable in characterizing the interactions between small molecules and their protein targets, typically those directed at cysteine residues.

Evotec has developed high-throughput mass spectrometry workflows to identify covalent binders for protein targets. Using both, Intact RF/MS and RF/MS-based peptide attenuation approaches, we screened Evotec’s 5k covalent library set comprising of weakly reactive acrylamide-based compounds to identify specific cysteine modifiers near the active site of an enzyme. Performance and sensitivity of both MS-based methods are presented and discussed.

Improving and Accelerating Hit Identification using High Throughput Chemistry (HTC) combined with Direct to Biology (D2B)

^{Date and Time: Wednesday, May 21, 2025: 4:30 PM–5:30 PM CEST (Presentation Number: 1146-A)
Location: CCH – Congress Center Hamburg – Exhibition Hall
Presented by: Pierre Zindy, Senior Research Scientist}

Plate-based High Throughput Chemistry (HTC) is combined with Direct to Biology (D2B) to prepare extensive compound libraries on a microscale, without purification. The crude mixtures are composed of the desired target molecules and are assessed in bioassays on the High Throughput Screening platforms. Selected compounds are then re-synthesized, purified, and validated in the same bioassays. Evotec is presenting its HTC/D2B workflow, including the “innocuity set”, to offer faster and optimized Structure Activity Relationship (SAR) exploration, that could take place at the Hit expansion, Hit-to-Lead, and Lead optimization phases. The poster illustrates some applications, taking advantage of successful implementations, implicating close collaboration between chemistry, high-throughput screening, and sample management teams.

Integrated Surface Plasmon Resonance Platform for Biologics Discovery Support

^{Date and Time: Wednesday, May 21, 2025: 4:30 PM–5:30 PM CEST (Presentation Number: 1150-A)
Location: CCH – Congress Center Hamburg – Exhibition Hall
Presented by: Florian Krieger, VP Biophysics}

Biologics have revolutionized disease treatment by overcoming the limitations of traditional small-molecule drugs. Evotec provides an end-to-end, flexible, and efficient antibody development platform - from discovery to clinical stages - ensuring rapid candidate selection and high-quality biologics. Surface Plasmon Resonance (SPR) is integral throughout the process, enabling detailed antibody characterization from off-rate ranking to Epitope Binning and Fc receptor binding. Evotec supports this with a standardized workflow and extensive SPR infrastructure, including multiple 8K+ and T200 devices across its sites.

Keeping an Eye on the Off Rate – Analysis of Interactions by SPR

^{Date and Time: Wednesday, May 21, 2025: 4:30 PM–5:30 PM CEST (Presentation Number: 1154-A)
Location: CCH – Congress Center Hamburg – Exhibition Hall
Presented by: Katrin Schaefer, Senior Research Scientist, Biophysics}

Surface plasmon resonance (SPR) is a powerful tool for studying molecular interactions across various molecular weights and affinities. It enables drug discovery by linking binding events to candidate potency, with kinetic parameters guiding lead selection. The off rate (kd) is crucial for experiment setup and interaction analysis, requiring different approaches for low-affinity fragments versus high-affinity protein-protein interactions. This study explores experimental workflows for diverse affinities, including fragment screening, hit-to-lead development, and chaser experiments to determine binding affinity.

DIY and Robotics: A Custom Integrated Cleaning System to Preserve Accuracy and Lifespan of Dispensers’ Cassettes on a HTS Platform

^{Date and Time: Wednesday, May 21, 2025: 4:30 PM–5:30 PM CEST (Presentation Number: 1162-A)
Location: CCH – Congress Center Hamburg – Exhibition Hall
Presented by: Jean-Philippe Navarro, Research Scientist, High Throughput Biology & Screening}

High-Throughput Screening (HTS) benefits from miniaturization, but low-volume dispensers require careful maintenance to prevent clogging. No manufacturers provide an automated cleaning system, so researchers at Evotec developed a custom autonomous cassette-washing system integrated on a HTS platform. It uses electro-valves to regulate cleaning liquids through the dispenser tubing, ensuring optimal performance and extending cassette lifespan. This innovation reduces consumable waste, eliminates manual cleaning, and enhances efficiency in continuous screening assays. The poster details the system's design, integration, and impact on HTS workflows.

Generating Lead-like Small Molecules Targeting a Structured 5’UTR mRNA Element in an APOBEC3B Synthetic Lethal Partner for Cancer Therapy

^{Date and Time: Wednesday, May 21, 2025: 4:30 PM–5:30 PM CEST (Presentation Number: 1168-A)
Location: CCH – Congress Center Hamburg – Exhibition Hall
Presented by: Steffen Grimm, VP RNA Biology}

The APOBEC3B mutational signature is prevalent in many tumors, offering both an advantage to cancer cells and a therapeutic vulnerability. Using CRISPR screening in triple-negative breast cancer, we identified synthetic lethal partners of APOBEC3B, including a target deemed "undruggable" by conventional approaches. Through our RNA-small molecule drug discovery platform, we found a druggable structured 5’UTR RNA element that could be selectively bound by optimized small molecules. Through SHAPE-MaP, AS-MS, SAXS, and molecular modeling, small molecule hits that bind this RNA element were identified and optimized, leading to selective target protein reduction in cells. This study highlights the potential of RNA-targeting small molecules to drug challenging cancer targets.

Sensitive Immunoassays Across Diverse Matrices: Enabling Minimally Invasive Biomarker Assessments

^{Date and Time: Wednesday, May 21, 2025: 4:30 PM–5:30 PM CEST (Presentation Number: 1174-A)
Location: CCH – Congress Center Hamburg – Exhibition Hall
Presented by: Alexander Weiss, Group Leader, Translational Biomarkers}

Evotec leverages ultra-sensitive immunoassays and minimally invasive sampling to improve early diagnosis, patient stratification and monitoring of complex diseases. Using platforms like Single Molecule Counting (SMC), Meso Scale Discovery (MSD), Quanterix, and NULISA, biomarkers are detected in fluids such as plasma, CSF, tears, and extracellular vesicles. Highlights include detecting mutant huntingtin in tears for Huntington’s disease and mutant ATXN3 in plasma/CSF for spinocerebellar ataxia type 3 (SCA3), as well as blood-based inflammatory markers and EV biomarker profiling for broader research use.

Discovery of Small Molecule Antivirals Targeting Hepatitis B Virus Epsilon Element

^{Date and Time: Thursday, May 22, 2025: 2:00 PM–3:00 PM CEST (Presentation Number: 1013-B)
Location: CCH – Congress Center Hamburg – Hall H 1013-B
Presented by: Steffen Grimm, VP RNA Biology}

The hepatitis B virus (HBV) epsilon (ε) RNA element is critical for viral replication and packaging and so is a key target for antiviral therapy. Using affinity selection mass spectrometry (AS-MS),  ~50,000 lead-like compounds were screened for binding to ε, with 121 active compounds identified. Of these, 75 were tested for antiviral activity in HepaD38 cells. Structure-relationship (SAR) studies led to the discovery of a promising chemical series with enhanced antiviral properties and desirable physicochemical properties. Analogues with improved profiles significantly reduced viral RNA and protein levels in HBV-infected cells with kinome, transcriptome and proteome analysis suggesting promising selectivity and specificity Future work will optimize these leads and explore their mode-of-action.

CRISPR-Walking Identifies mRNA-Regulatory Elements within the Untranslated Regions of Target Genes

^{Date and Time: Thursday, May 22, 2025: 2:00 PM–3:00 PM CEST (Presentation Number: 1111-B)
Location: CCH – Congress Center Hamburg – Hall H
Presented by: Melanie Winkle, Research Scientist, In Vitro Biology}

CRISPR-walking identifies mRNA-regulatory elements within untranslated regions (UTRs) by systematically screening for sequences that affect mRNA stability or translation, offering a potential new route for therapeutic targeting. In this study, CRISPR-walking was used to tile the entire 3'UTR of Target X with 152 gRNAs in HeLa cells. The screen successfully pinpointed two known regulatory elements - confirmed by increased protein expression (2-4.3x) - and revealed a novel region with potential repressive function. The method proved robust, with >80% editing efficiency and consistent results across various gRNA types. This approach offers a powerful tool for identifying mRNA-regulatory elements and advancing RNA-targeted drug development.

Increased Efficiency for Ion Channel Drug Discovery: Case Studies from a Screening Campaign on the Automated Patch Clamp Platform Qube

^{Date and Time: Thursday, May 22, 2025: 2:00 PM–3:00 PM CEST (Presentation Number: 1147-B)
Location: CCH – Congress Center Hamburg – Hall H 
Presented by: Nasreen Choudhury, Research Scientist, In Vitro Biology}

Ion channels represent a significant target for drug discovery, with 18% of approved small-molecule drugs acting on them. High Throughput Screening (HTS) using automated patch clamp (APC) electrophysiology platforms provides direct and specific results but often comes with longer timelines and higher costs compared to indirect fluorescence-based methods.

At Evotec, efficiency in ion channel HTS was improved using the APC Qube platform in two ways:

1. Unattended Runs - Allowing automated operation without constant supervision, reducing the campaign timeline from 7 months to 3.5 months while maintaining high screening quality.
2. Assay Ready Frozen (ARF) Inducible Cells - Optimizing cell preparation to streamline off-target compound testing, reducing operational time and cost.

These adaptations enhanced automation, efficiency, and cost-effectiveness in drug discovery, though each APC assay and cell line requires individual optimization.

Integration of Acoustic Ejection Mass Spectrometry for HTS-based Hit Identification Demonstrated on SARS-CoV2 Mpro

^{Date and Time: Thursday, May 22, 2025: 2:00 PM–3:00 PM CEST (Presentation Number: 1151-B)
Location: CCH – Congress Center Hamburg – Hall H
Presented by: Florian-Alexander Herbst, Group Leader, Mass Spectrometry}

This study demonstrates the successful integration of Acoustic Ejection Mass Spectrometry (AEMS) into high-throughput screening (HTS) workflows, using SARS-CoV-2 main protease (Mpro) as a model target. AEMS enables label-free mass spectrometry at optical-readout speeds. Through integration with a Beckman access workstation, a biochemical assay for Mpro was developed and automated for HTS, screening 50k diverse lead-like compounds and 5k covalent compounds across Evotec’s libraries. Hits were confirmed and profiled for IC₅₀ values. The pilot screen confirmed assay performance and sensitivity, validating AEMS as a reliable HTS readout method.

Leveraging Sample Management Strengths to Support Biologics Research

^{Date and Time: Thursday, May 22, 2025: 2:00 PM–3:00 PM CEST (Presentation Number: 1155-B)
Location: CCH – Congress Center Hamburg – Hall H
Presented by: Bastian Pfeiffer, Research Associate, Compound Management}

With the increasing focus on biologics in drug discovery, Evotec’s sample management teams have developed reliable water-based workflows to support internal research. This study highlights the use of the Beckman Coulter Echo 655T for precise nanoliter dispensing, detailing the quality control measures that ensure consistent performance. Key challenges such as source plate evaporation and concentration stability are addressed, along with the strategies implemented to mitigate them. These optimized processes have successfully enabled the CRISPR team to miniaturize assays, achieving cost savings while maintaining data quality.

Optimizing 1D ¹³C NMR Spectroscopy for Protein Secondary Structure Prediction: Application on a TCI Micro-CryoProbe Head using Natural Abundance ¹³C Samples

^{Date and Time: Thursday, May 22, 2025: 2:00 PM–3:00 PM CEST (Presentation Number: 1165-B)
Location: CCH – Congress Center Hamburg – Hall H
Presented by: Jan Kahmann, Principal Scientist Biophysics - NMR}

This study optimizes 1D ¹³C NMR spectroscopy for predicting protein secondary structures using natural abundance ¹³C and a TCI Micro-CryoProbe, typically optimized for ¹H detection. Using the SSD-NMR method and a specialized experiment ("zgig_pisp") to reduce baseline distortion, alongside Cu(II) EDTA to accelerate scan times, the team successfully acquired usable spectra from low-volume (50 µL) samples in under 24 hours. The resulting data allowed proper secondary structure predictions, supported by a tailored data processing pipeline. Future work may include testing 10% ¹³C-spiked samples to further reduce acquisition times.

Phenotypic Screening – Application of Cell Painting to Accelerate Drug Discovery

^{Date and Time: Thursday, May 22, 2025: 2:00 PM–3:00 PM CEST (Presentation Number: 1169-B)
Location: CCH – Congress Center Hamburg – Hall H
Presented by: Sven Schreiter, Senior Research Scientist, High Throughput Biology}

Evotec uses high content imaging to support drug discovery, enabling analysis of small molecule effects on multiple cellular processes. A key approach, Cell Painting, supports phenotype characterization, hit identification, target deconvolution, mode-of-action prediction, and early safety assessment. Enhancements like well randomization and miniaturization to 1536-well plates have improved data quality and throughput. These robust datasets feed AI/ML models for optimized hit selection.

The Future of Drug Discovery: Connecting Biophysical Data with AI/ML Methods

^{Date and Time: Wednesday, May 21, 2025: 12:30 PM–1:00 PM CEST
Location: CCH – Congress Center Hamburg – Hall F
Presented by: Florian Krieger, VP Biophysics}

AI-based tools in drug discovery are rapidly advancing. Recent examples include AlphaFold-based tools for accurate protein structure prediction, or the combination of high throughput screening (HTS) with machine learning (ML) for optimized hit enrichment. Despite these advances, few clinical candidates have been identified from computational approaches due to limited relevant and high quality data sets. To improve the translation of AI/ML models into physiologically relevant contexts, multiple software tools and biophysical experiments are being evaluated. These tools simulate 1D ¹³C NMR-spectra and correlate experimental and in silico parameters. Additionally, ML-based hit enrichment steps are being implemented to accelerate the transition from fragment hits to lead-like chemical space using biophysical readouts like SPR or NMR.

Scalable Genetic Tools for High Throughput Target Validation and Deconvolution

^{Date and Time: Wednesday, May 21, 2025: 12:30 PM–1:00 PM CEST
Location: CCH – Congress Center Hamburg – Hall D
Presented by: Hauke Cornils, VP Early Discovery}

Evotec has integrated CRISPR/Cas9 technology into its drug discovery platform, enabling large-scale functional genomics screening for target identification and validation. In this presentation, three case studies are presented:

1. Application of CRISPR screening as a tool for high-throughput target validation in the context of synthetic lethality.
2. Target validation of 300 genes from diabetic kidney disease data.
3. Genetic approaches to understand compound activity.

These case studies highlight the power of CRISPR technology in the discovery of novel disease relevant targets.

Covalent High-Throughput Mass Spectrometry: Tough Targets Beware!

^{Date and Time: Wednesday, May 21, 2025: 1:00–2:00 PM CEST
Location: CCH – Congress Center Hamburg – Hall B03-04
Presented by: Florian-Alexander Herbst , Group Leader, Mass Spectrometry, HTS and Julien Parra, Senior Research Scientist, HT Biology and Screening}

Covalent inhibitors are transforming drug discovery with success in hitting targets which were previously considered undruggable. Evotec uses High-Throughput Mass Spectrometry (MS) workflows for covalent screening and rapid hit identification. This presentation highlights the important factors in designing a covalent library, how high throughput MS is used to analyze covalent modifications and how leads are validated in a cellular setting.

SmartCel-2 Coming to the Rescue: How We Overcame Capacity Challenges with Existing Equipment

^{Date and Time: Wednesday, May 21, 2025: 3:30–4:00 PM CEST
Location: CCH – Congress Center Hamburg – Hall E
Presented by: Isabelle Bergere, VP Head of Sample Management}

In response to a doubling in serial dilution preparations over the past five years, we implemented the SmartCel-2 automation system from Flexible Lab Solutions to meet the increased demand. This presentation outlines our collaboration with the provider, from design to implementation, tailored to our specific needs. We will discuss the selection of instruments, quality maintenance strategies, and the user interface that accommodates process variabilities. Additionally, we will highlight the smart scheduling software that enhances efficiency and throughput.

High Dimensional Cell Painting Analysis Applied Across All Drug Discovery Phases

^{Date and Time: Thursday, May 22, 2025: 12:30–1:00 PM CEST
Location: CCH – Congress Center Hamburg – Hall D
Presented by: Karsten Kottig, VP Imaging & Analysis}

The shift from traditional image analysis to high-dimensional readouts with predictive analytics has transformed data handling and interpretation. Cell Painting, a large-scale phenotypic fingerprinting technique, enables the integration of diverse data platforms.
The presentation highlights imaging and data analysis applications in the drug discovery process, showcasing the transition to high-dimensional data interpretation and predictive data science. Examples presented include target validation and deconvolution, hit identification, and early safety/toxicity prediction, demonstrating the benefits of modern data generation and analysis techniques.

Drugging the Undruggable - Mass Spectrometry-Based Proteomics Screening at Industrial Scale

^{Date and Time: Thursday, May 22, 2025: 3:00 PM–3:30 PM CEST
Location: CCH – Congress Center Hamburg – Hall D
Presented by: Christoph Schaab, SVP Head of Proteomics & Metabolomics}

Over the past two decades, new strategies like targeted protein degraders, stabilizers, and covalent ligands have emerged to target previously undruggable proteins. Proteomic technologies, particularly LC-MS-based quantitative proteomics, have revolutionized the discovery process from hit identification to preclinical development.

In the presentation, advanced workflows and instrumentation for proteome-scale profiling (ScreenPep™) are discussed including the identification of targets degraded by molecular glues and the validation of their mode-of-action. Additionally, high-throughput activity-based protein profiling (HT-ABPP) maps reactive cysteine sites and screens fragment libraries to identify hits. These platforms, along with HT-PALMS and intact protein MS, provide multiple routes for discovering new therapeutic targets and compounds.