4) Procedure

The following steps outline the detailed procedure for handling HTS equipment:

1. Step 1: Preparation of HTS Equipment
   1. Ensure that the HTS equipment is set up correctly before use. Verify that all components (e.g., robotic arms, plate readers, liquid handling systems) are properly installed and connected.
   2. Check the consumables (e.g., assay plates, tips, reagents) for quality and ensure sufficient quantities are available for the screening process.
   3. Calibrate the equipment, including plate readers and liquid handlers, according to the manufacturer’s specifications. Calibration should be performed regularly to maintain accuracy and precision in measurements.
   4. Conduct a dry run of the equipment to ensure that all components are functioning as expected before loading any compounds or samples.
2. Step 2: Loading Samples and Compounds
   1. Prepare the samples and compounds to be screened. Ensure that all samples are properly labeled and stored according to the required conditions (e.g., temperature, humidity).
   2. Load the samples or compounds into the HTS system, ensuring that they are placed in the appropriate well plates. Ensure that each sample is accurately pipetted to prevent cross-contamination between samples.
   3. Double-check that the sample plates are correctly aligned and positioned within the HTS system. Verify that no wells are missed or improperly loaded.
3. Step 3: Running the Screening Process
   1. Start the screening process according to the established protocol. Ensure that the correct assay conditions (e.g., temperature, time, reagent volumes) are applied for each specific screen.
   2. Monitor the screening process to ensure that the equipment is operating without interruptions. Address any issues, such as sample delivery failures or malfunctioning robotic arms, immediately to avoid delays.
   3. During screening, ensure that all safety measures are followed to protect operators and the integrity of the samples (e.g., proper handling of hazardous materials or biological agents).
4. Step 4: Data Collection and Analysis
   1. Once the screening run is complete, collect the data generated by the HTS equipment, including assay readouts (e.g., fluorescence, luminescence, absorbance) and compound screening results.
   2. Verify that all data is captured correctly and stored in the appropriate data management systems. Ensure that the data is formatted for easy analysis and that it complies with data integrity standards.
   3. Perform initial analysis of the screening results to identify potential hits or lead compounds. This can include hit validation using secondary assays or data visualization tools.
5. Step 5: Equipment Maintenance and Troubleshooting
   1. Perform routine maintenance on the HTS equipment, including cleaning the robotic arms, replacing consumables (e.g., tips, plates), and verifying fluidics systems are functioning properly.
   2. Calibrate the equipment at regular intervals to ensure continued accuracy. Perform any necessary adjustments to settings based on calibration results.
   3. If any issues arise during the screening process (e.g., mechanical failure, inaccurate readings), troubleshoot the equipment promptly. Refer to the equipment’s user manual or contact technical support if the issue cannot be resolved in-house.
6. Step 6: Documentation and Reporting
   1. Document all procedures, including equipment setup, sample loading, screening protocols, calibration records, and maintenance activities.
   2. Ensure that all data collected from HTS runs are properly recorded and stored. Generate reports summarizing the screening results and any technical issues encountered during the process.
   3. Ensure that all HTS activities are fully compliant with internal protocols, regulatory standards, and documentation requirements for regulatory filings, if applicable.

5) Abbreviations

• HTS: High-Throughput Screening
• QA: Quality Assurance
• ADMET: Absorption, Distribution, Metabolism, Excretion, Toxicity
• RPM: Revolutions Per Minute (related to liquid handling speed)
• QC: Quality Control

6) Documents

The following documents should be maintained throughout the HTS process:

1. HTS Screening Protocols
2. Equipment Calibration and Maintenance Logs
3. Screening Data and Results Reports
4. Sample Loading Records
5. Troubleshooting and Incident Reports

7) Reference

References to regulatory guidelines and scientific literature that support this SOP:

• FDA Guidance for Industry on Drug Discovery and Screening
• Standard protocols for High-Throughput Screening (HTS)
• Scientific literature on the best practices for HTS equipment operation and maintenance

8) SOP Version

Version 1.0: Initial version of the SOP.