Drug discovery campaigns need not be limited to strategies with focus on single or strictly defined targets. Instead, target-agnostic programmes utilizing validated phenotypic criteria to detect promising therapy-like activities can deliver candidate hit molecules without underlying assumptions regarding target identities.
At o2h discovery, we utilize the CellInsight CX7 LED Pro HCS platform, allowing our skilled Biology team to tailor phenotypic features to meet the specific requirements of your target biology assays. By applying algorithmic image analysis principles, we can quantitatively assess these features, supporting a diverse range of high-content imaging assay formats for your drug discovery projects.
Case Study 1:
o2h discovery specializes in developing cellular assays that integrate biological reporter systems, allowing for quantitative assessment of target engagement. These assays are invaluable for structure-activity relationship (SAR) studies, demonstrating dose-dependency, and validating cell-free predictions of binding potencies to differentiate candidate molecules effectively.
Our HCI phenotypic assays, utilizing stable cell lines with tagged reporters, are particularly advantageous in scenarios where scale, cost-per-well, and rapid turnover are critical factors in the drug discovery screening cycle. This approach ensures efficient and informative screening, enabling more informed decision-making in your drug development process.
In the above provided example, the raw data from the HCI assay tracks the sub-cellular translocation of a GFP-tagged reporter in response to either vehicle treatment or an experimental small molecule.
Above, you can see the HCI nuclear reporter translocation data, which serves as a dose-dependent phenotypic measure of cellular target engagement. This analysis allows us to quantify how the reporter’s localization changes with varying concentrations of the small molecule, providing insights into its mechanism of action and efficacy. Such data are crucial for understanding the dynamics of target engagement and optimizing lead compounds in the drug discovery process.
Case Study 2:
In support of drug discovery campaigns, the HCI experts at o2h discovery can create assays that track cell phenotypes indicative of target engagement. One notable example is our HCI speckle assay, which detects cellular activity by monitoring the punctate versus diffuse states of endogenous spliceosome factors. In this assay, small molecules are evaluated for their ability to induce dose-dependent phenotypic shifts, reflecting their engagement with a target kinase known to regulate mRNA splicing. This approach not only provides insights into the mechanisms of action of potential drug candidates but also helps identify compounds that effectively modulate cellular processes related to splicing
In the above example HCI data in which MCF7 cells were treated with either vehicle or a tool compound and subsequently stained with fluorescent markers, enabling algorithmic detection and characterization of spliceosome speckles
In the above HCI speckle assay performed in MCF7 cells, we generate dose-response curves guiding structure-activity relationship (SAR) development
Case Study 3:
Assay development on HCI systems facilitates innovative approaches to information management and study design. At o2h discovery, we integrated HCI assay development with the SemaCyte® cell microcarrier platform developed by Semarion.
Combining HCI with this microcarrier system enables novel strategies for managing cell monolayers in drug discovery workflows. Key enhancements include the miniaturization of cell seeding, freezing of assay-ready adherent cells, and multiplexing of cell lines using subpopulation barcoding.
Powered by HCI image analysis, this methodology provides significant benefits for projects that require economical cell seeding, efficient utilization of premium assay reagents, and advanced internal control strategies.
To know more about our biology services offering or to request our brochure, please reach out to us at discovery@o2h.com.
