Assay Development (Lake Nona)

Overview

The high-throughput screening (HTS) assay development team provides support for the design and development of new HTS assays as well as optimization support for existing assays. In Lake Nona in Orlando, the facility shares space with the ultra-high-throughput (uHT) screening center which houses the modular HighRes screening system instrumentation. The system may be used in workstation mode by the facility team for seemless implementation of assays within the fully integrated system together with the uHTS team. The assay development team is skilled in various screening modalities, to help with selection and development of assays most suitable to each particular project. The team also provides support in the design of optimal structure-activity relationship (SAR) panels, development of supporting assays, data analysis, and interpretation for studies aimed at characterization and optimization of hits obtained in primary HTS.

The facility is a part of Conrad Prebys Center for Chemical Genomics and can advise and help in preparation of data packages for screening-related grants.
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Services

  • Consultations on suitable screening strategies and alternatives:
    We provide consultations on (1) developing primary screening platforms for uHTS; (2) designing a screening cascade including secondary and tertiary assays to maximize the success of identifying a potent, selective, and chemically tractable small molecule probe; and (3) preparing screening-related grant applications.
  • Assay design and development:
    We develop biochemical and cell-based assays (including phenotypic and high-content assays) using commercial technologies. To validate assay robustness and performance in 384-wells, we may test against the library collection of 1,280 pharmacologically active and commercially available compounds (LOPAC).
  • Implementation and optimization of previously published screening protocols:
    We miniaturize and optimize assays into 1536-wells. Z'-factors are determined from both negative and positive controls and are used to evaluate assay reproducibility, stability, and scalabilty. We test DMSO tolerability and test multiple plates containing DMSO (compound vehicle) to establish false positive and false negative frequencies. Assays that may be tested against one or more compound library collections.
  • Support for structure-activity relationship studies:
    Once the HTS campaign has been completed, a dedicated chemical biology team comprised of assay development support, cheminformatics, and medicinal chemistry confirms the hit set in dose-response in the primary assay and develops supporting assays to prioritize scaffolds from the validated hit set, and support SAR activities.
  • Project management:
    The project team at Lake Nona in Orlando collaborates with principal investigators to establish a comprehensive project plan and review project roles, responsible parties, and confirm or revise timelines and milestones.

Equipment and Resources

Robotic equipment and resources are used by both the assay development team as well as the uHTS facility in Lake Nona in Orlando, and they constitute a very extensive pool of technology. Click here for a complete and detailed list or our resources.
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