Addressing Unusual Assay Variability with Robust Statistics

Well-behaved, in vitro bioassays generally produce normally distributed values in their primary (efficacy) data. Accordingly, the best practices for statistical analysis are well documented and understood. However, assays may occasionally display unusually high variability and fall outside the assumptions inherent in these standard analyses. These assays may still be in the optimization phase, in which the source of variation could be identified and addressed. They might also represent the best available option to address the biological process being examined. In these cases, the use of robust statistical methods may provide a more appropriate set of tools for both data analysis and assay optimization. This article provides guidance on best practices for the use of robust statistical methods for the analysis of bioassay data as an alternative to standard methods. Impacts on experimental design and interpretation will be discussed.

1. Iversen, P. W., Beck, B., Chen, Y. F., et al. HTS assay validation. In Assay Guidance Manual; Markossian, S., Sittampalam, G. S., Grossman, A., et al., Eds.; 2012.
https://www.ncbi.nlm.nih.gov/books/NBK83783/
Google Scholar2. Beck, B., Chen, Y. F., Dere, W., et al. Assay operations for SAR support. In Assay Guidance Manual; Markossian, S., Sittampalam, G. S., Grossman, A., et al., Eds.; 2012. https://www.ncbi.nlm.nih.gov/books/NBK91994/
Google Scholar3. Haas, J. V., Eastwood, B. J., Iversen, P. W., et al. Minimum significant ratio—a statistic to assess assay variability. In Assay Guidance Manual; Markossian, S., Sittampalam, G. S., Grossman, A., et al., Eds.; 2013. https://www.ncbi.nlm.nih.gov/books/NBK169432/
Google Scholar4. Eastwood, B. J., Farmen, M. W., Iversen, P. W., et al. The minimum significant ratio: a statistical parameter to characterize the reproducibility of potency estimates from concentration-response assays and estimation by replicate-experiment studies. J. Biomol. Screen. 2006, 11, 253–261.
Google Scholar | SAGE Journals5. Huber, P. J., Ronchetti, E. M. Robust Statistics; 2nd ed.; Wiley: New York, 2009.
Google Scholar | Crossref6. Hanson, J. D., Condroski, K., Correaet, M., et al. Protein Degradation via CRL4CRBN Ubiquitin Ligase: Discovery and Structure–Activity Relationships of Novel Glutarimide Analogs That Promote Degradation of Aiolos and/or GSPT1. J. Med. Chem. 2018, 61, 492–503.
Google Scholar | Crossref | Medline7. Zhang, J. H., Chung, T. D. Y., Oldenburg, K. R. A Simple Statistical Parameter for Use in Evaluation and Validation of High Throughput Screening Assays. J. Biomol. Screen. 1999, 4, 67–73.
Google Scholar | SAGE Journals8. McGill, R., Tukey, J. W., Larsen, W. A. Variations of Box Plots. Am. Stat. 1978, 32, 12–16.
Google Scholar

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