How Automation Overcomes Variability & Limitations in HTS Troubleshooting

High-throughput screening (HTS) is an integral part of the drug discovery workflow, allowing the rapid testing of large compound libraries to identify potential therapeutic candidates¹. However, researchers regularly encounter frustrating limitations, particularly in HTS troubleshooting². In this article, we will discuss the most common challenges in HTS troubleshooting, explore how automated technologies can help researchers overcome them, and outline strategies for successful automation implementation. 

Challenges in HTS Troubleshooting

HTS is used extensively in drug discovery to test the effect of numerous compounds on a target simultaneously. However, HTS can produce false positives or false negatives, which may lead to wasted resources or missed opportunities, respectively^2,3. Addressing these issues requires effective HTS troubleshooting. Some common challenges in HTS troubleshooting include: 

• Variability. Like most laboratory workflows, HTS consists of manual processes, which are subject to inter- and intra-user variability. Even tiny variations between how two different users perform processes can lead to huge discrepancies in results, as exemplified by over 70% of researchers reporting being unable to reproduce the work of others⁴! The lack of standardization in processes can make HTS troubleshooting difficult. 

• Human error. Manual processes are also subject to human error, which can lead to inconsistencies and unreliable results. Since errors often go unnoticed, they also frequently remain undocumented, which makes HTS troubleshooting even more of a minefield. 

• Data handling challenges. HTS produces vast volumes of multiparametric data, which can be challenging to manage, store, and analyze effectively to gain meaningful insights and accurately identify hit compounds⁵. 

• Logistical difficulties. Establishing effective HTS procedures requires careful selection of equipment, design of experiments, validation of processes, and development of robust data analysis methodologies, which can present numerous obstacles. 

The Role of Automation in HTS

Automation is an essential component of HTS workflows; it enables researchers to streamline workflows and overcome some of the hurdles associated with HTS and HTS troubleshooting. Automation has several key benefits in HTS troubleshooting: 

• Enhanced data quality and reproducibility. Automated workflows improve assay performance and reproducibility, standardizing workflows and reducing variability and errors across users, assays, and sites⁶. Advanced tools further improve data reliability by providing in-built verification features. For example, the I.DOT Liquid Handler is equipped with DropDetection technology, which verifies that the correct volume of liquid has been dispensed into each well. This supports HTS troubleshooting by allowing dispensing errors to be identified, documented, and corrected. 

• Increased throughput and efficiency. Automated systems enable large compound libraries to be screened at multiple concentrations, generating comprehensive data sets and significantly increasing throughput⁷. This can be very useful in HTS troubleshooting, where many conditions must be tested to identify the source of error or optimal conditions. 

• Scalability and flexibility. Automated platforms allow for the development and optimization of flexible HTS protocols, which can be adapted and scaled as and when project needs require. 

• Cost reduction. HTS involves screening many samples, which uses vast amounts of resources. Automation enables miniaturization, reducing reagent consumption and overall costs by up to 90%⁸. This makes it feasible to conduct comprehensive analyses even with limited samples and cuts costs substantially. 

• Data management. In addition to the bench-based workflow, data management and analytical processes can be automated, streamlining the process and enabling rapid insights in HTS troubleshooting and workflows for faster drug development pipelines (Fig. 1)⁹. 

Figure 1. Automated data management and analytics enable streamlined analysis of complex HTS data, supporting faster time to insights. (Source)

Implementing Automation in HTS Workflows

Integrating automation into HTS troubleshooting and workflows can significantly improve efficiency, accuracy, and reproducibility, but careful planning is necessary for a successful transition. First, current workflows should be assessed to identify the bottlenecks and labor-intensive tasks that could benefit from automation, such as liquid handling, compound dilutions, and data analysis, where automation can reduce human error and increase throughput.

When selecting automation tools, the lab’s specific requirements in terms of scale and workflow flexibility must be considered. For example, liquid handling tools like the I.DOT Non-Contact Dispenser excel in offering high precision at low volumes. Meanwhile, robotic arms and integrated systems that can handle multiple plates at once might be more appropriate for larger-scale screening, and the I.DOT Non-Contact Dispenser has been designed to be integrated into any automated work cell (Fig. 2). Additionally, assessing the technical support, ease of use, and software integration of potential technologies is essential to ensure a smooth transition.

Figure 2. The I.DOT Liquid Handler can be seamlessly and efficiently incorporated into any automated solution to maximize efficiency, reproducibility, and process safety.

Conclusion

Automation plays a critical role in overcoming the challenges and limitations of HTS, particularly in HTS troubleshooting. By reducing human error, increasing reproducibility, and enhancing data quality, automated systems streamline HTS workflows and minimize variability. Tools like the I.DOT Liquid Handler offer high precision, scalability, and flexibility, enabling researchers to troubleshoot HTS efficiently while reducing costs and turnaround times. As automation continues to advance, its integration into HTS workflows will be essential for optimizing drug discovery processes and ensuring accurate, reliable results.

Ready to Revolutionize Your HTS Process?

Discover how the I.DOT can eliminate variability and enhance your screening accuracy. Download the I.DOT brochure to learn more about the precision and efficiency that the I.DOT brings to your lab!

References

1. Aherne GW, McDonald E, Workman P. Finding the needle in the haystack: why high-throughput screening is good for your health. Breast Cancer Res. 2002;4(4):148. doi:10.1186/bcr440
2. Sink R, Gobec S, Pecar S, Zega A. False Positives in the Early Stages of Drug Discovery. Curr Med Chem. 2010;17(34):4231-4255. doi:10.2174/092986710793348545
3. Burt T, Button K, Thom H, Noveck R, Munafò M. The Burden of the “False‐Negatives” in Clinical Development: Analyses of Current and Alternative Scenarios and Corrective Measures. Clin Transl Sci. 2017;10(6):470-479. doi:10.1111/cts.12478
4. Baker M. 1,500 scientists lift the lid on reproducibility. Nature. 2016;533(7604):452-454. doi:10.1038/533452a
5. Major J. Challenges and Opportunities in High Throughput Screening: Implications for New Technologies. SLAS Discov. 1998;3(1):13-17. doi:10.1177/108705719800300102
6. Holland I, Davies JA. Automation in the Life Science Research Laboratory. Front Bioeng Biotechnol. 2020;8:571777. doi:10.3389/fbioe.2020.571777
7. Michael S, Auld D, Klumpp C, et al. A Robotic Platform for Quantitative High-Throughput Screening. ASSAY Drug Dev Technol. 2008;6(5):637-657. doi:10.1089/adt.2008.150
8. Browne DJ, Kelly AM, Brady JL, Doolan DL. A high-throughput screening RT-qPCR assay for quantifying surrogate markers of immunity from PBMCs. Front Immunol. 2022;13:962220. doi:10.3389/fimmu.2022.962220
9. Nicolaou C. Automated Lead Discovery and Development in HTS Datasets. J Assoc Lab Autom. 2001;6(2):60-62. doi:10.1016/S1535-5535(04)00129-7