High Throughput Screening

Sygnature Discovery is a leading Contract Research Organisation (CRO) offering High Throughput Screening along with other hit identification tools such as Virtual Screening, Knowledge-based design, and fragment screening. Offering an integrated strategy for hit identification, we apply the most appropriate approaches to discover hit series that have the highest potential for developing into compounds with drug-like properties.

High Throughput Screening, is a recognised method in the drug discovery sector, swiftly assists in finding new chemical beginnings by integrating medicinal chemistry, biology, informatics, and state-of-the-art automation technologies.

Due to the complexities of this process, many factors outside of the core assay biology influence the quality and outcome of a High Throughput Screen. These include the quality of the library and strategies for identifying hits. Our HTS libraries and capabilities were developed to address these complexities, ensuring the quality of the hits and the data we provide across all target classes and assay formats.

The HTS group at Sygnature have a wealth of experience running HTSs across a wide range of target classes, therapeutic areas, and assay technologies.

There are many components involved in generating high-quality starting points for a drug discovery project. These include quality robust assays, reliable automation, secure data, and the quality of the libraries. At Sygnature Discovery, we meticulously factor in all these elements, both in the creation of our High Throughput Screening libraries capabilities and throughout each project, to ensure optimal results for our customers.

• Compound Libraries
• HTS Workflow
• Compound tracking and storage
• Automation
• Data analysis and fidelity

Compound Libraries

Our High Throughput Screening compound libraries are crucial for high-quality drug discovery starting points. Among these, our computational chemists have designed the LeadFinder Diversity Library. Additionally, we offer customisable High Throughput Screening by including the option to screen other commercial or customer-provided libraries.

LeadFinder Diversity Library (150k)

• Extensive Collection: 150,000 Compounds – Perfect for HTS Follow-Up.
• Expertly Designed: Low Molecular Weight, Lead-Like Properties.
• Diverse Structures: Strict Control on Similarity.
• Quality Assured: Each Compound inspected by Sygnature Discovery’s Senior Medicinal Chemists.
• Ready Resources: Freshly Sourced Solids, with Analogues and Building Blocks for Most Compounds.

LeadFinder Prism library (48k)

The Prism library, designed by Sygnature Discovery and developed for the European Lead Factory, contains 37,000 compounds. An extra 11,000 were internally produced using similar principles, with further compounds actively synthesised and added to the library annually.

• Compounds are drug-like with high chirality and FSP3
• Library contains novel natural product-inspired scaffolds

The Prism Library offers exclusive access to up to 50 hits per screen for enhanced intellectual property protection. We also ensure that Hit structures are excluded from subsequent screens for two years after the completion of each project.

Additional library sets:

• Enamine Protein-Protein Interaction library (40k) – lead-like compounds incorporating specific protein recognition patterns.
• Enamine Kinase Focused library (24k) – based on >100 hinge binding cores ((≥2 H-bonds possible with hinge)

If you wish to screen libraries in addition to these, we offer the option to screen customer or other commercial libraries.

High Throughput Screening Workflow

Each project is tailored to our customers’ requirements, with particular emphasis on developing robust quality assays. A key strength of the HTS group is our flexible approach, adapting each workflow as required. This is most evident when selecting hits to progress, where a balance is sought between false positive reduction and reducing the risk of including false negatives. Dedicated scientists along with a project coordinator are assigned to your project and are overseen by a senior member of the HTS team.

HTS Process:

• Begins with assay development or transfer.
• Once a robust, reliable assay has been established the validation (pilot) screen is performed.
• Pilot screen tests assay automation performance and give an early hit rate estimate.

Primary Screen:

• Typically designed to test a single concentration per compound from chosen libraries.
• Includes in-house and external libraries, plus any customer-provided libraries.

Hit Triaging and Selection:

• After the primary screen, hits undergo two cheminformatic triage stages.
• The first is to select compounds for hit confirmation and counter-screening.
• The second is to select compounds for final compound profiling with concentration-response curves.
  • The rigorous computational chemistry triage process in addition to targeted counter screens and orthogonal assays are run in collaboration with our customers team and highly skilled Sygnature Discovery computational chemists.

Hit Profiling:

• We take a nuanced and flexible approach to hit selection allowing for different levels of performance, hit rates and varying project requirements.
• This ensures we give the project the best chance of identifying quality hits
• Final compound profiling with concentration-response curves

Enhanced Compound Tracking and Storage at Sygnature Discovery

• Integrated Informatics:  Our High Throughput Screening (HTS) system uses a fully integrated informatics platform for superior data fidelity and precise compound tracking.
• Efficient Ordering: Utilising Titian Mosaic SampleBank software, linked with our HighRes® automated system, we streamline the ordering of assay plates.
• Optimal Storage Conditions: HTS libraries are stored in a custom-built facility with controlled low humidity and ambient temperature, ensuring compound integrity. HighRes® stackers organise HTS source plates for accessibility.
• Continuous Quality Monitoring: We employ continuous feedback from the HighRes® system to Mosaic, supplemented by regular library QC, to maintain compound quality.

Quality Control at Sygnature Discovery

• Quality Assurance through LCMS: To maintain the highest quality of compounds in our High Throughput Screening (HTS) campaigns, we regularly perform quality control (QC) checks. Using Liquid Chromatography-Mass Spectrometry (LCMS), we meticulously analyse samples from our HTS library source plates.
• Comprehensive QC in Hit Validation: Compounds that advance to the final stage of the hit validation phase, specifically during potency assessments (EC/IC50 determinations), undergo additional QC processes.

Integrated HTS and Compound Management:

• Our HTS group operates in close coordination with the Compound Management Department.
• This collaboration is enhanced by our automated Azenta Store, which facilitates the inclusion and management of external screening libraries.

Advanced Automation in Sygnature Discovery’s HTS System

• High Flexibility Design: Our High-Throughput Screening (HTS) system, developed by HighRes Biosolutions, is designed for versatility, accommodating both cell and biochemical assays.
• Precision Dispensing: Central to our automation is the Echo acoustic dispensing technology, ensuring highly accurate compound delivery.
• Reliability and Extended Operation: The system operates with remarkable reliability, routinely functioning for 20-30 hours continuously, and is available 24/7.
• Remote Monitoring and Control: Enhanced with full remote accessibility and CCTV monitoring, the system allows for prompt error detection and recovery.

Dynamic Scheduling for Efficiency:

Utilising Cellario for dynamic automation scheduling, our system ensures uniform treatment across assays, even for those challenging to schedule. Sygnature Discovery integrates cutting-edge automation to ensure efficiency and precision in our HTS processes.

Data Analysis Excellence in HTS at Sygnature Discovery

• Handling Complex Data: Our information technology system is specifically designed to manage and analyse large, complex datasets, a crucial capability for High-Throughput Screening (HTS).
• Genedata Screener Utilisation: We leverage Genedata Screener for data processing. This robust tool ensures efficient data management and analysis.
• Seamless Data Integration: Barcoded plate files are uploaded to Genedata Screener. The integration of plate barcodes with sample ID and location data from Mosaic guarantees data fidelity.
• Efficient Data Analysis: This system enables the rapid upload and thorough interrogation of screening data, facilitating detailed analysis by plate, batch, and screen.

Sygnature Discovery prioritises advanced data analysis capabilities to ensure accuracy and efficiency in HTS processes.