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Principal Investigator: Samuel Michael
Organization: NATIONAL CENTER FOR ADVANCING TRANSLATIONAL SCIENCES
Fiscal Year: 2020
Award: $10,097,450
Funding agency: National Center for Advancing Translational Sciences
Research Services Core (RSC)staff provide support to the NCATS activities in high-throughput screening, assay development, and optimization as well as follow-up and dose-response library-plating services for NCATS screening activities. The Compound Management team works in conjunction with the Automation team to provide follow-up and dose-response library-plating services for NCATS screening activities. Systems supported by the Automation team include primary screening system, Tox21 screening system, RNAi screening system, the RNAi hit picking system for assay follow ups, and the High Resolution Biosystems (HRB) acoustic plating system. In the past year, we have also installed and begun to operate a Thermo Scientific Spinnaker plate preparation workstation used to pin compounds and dispense reagents into plates in a batch format. Together, all of these systems combined provide a wide variety of automation and compound plating tasks used to enable all facets of the NCATS intramural scientific program.
The primary screening system utilizes three robotic arms for plate transportation, a storage capacity of more than 3 million compound wells (approximately 2,000 1,536-well compound plates), and more than 1.5 million assay wells (approximately 1,100 1,536-well assay plates) to achieve a world-class screening productivity while maintaining the high level of data quality required by researchers. It also has the ability to run multiple assays in parallel. Center experts have developed custom software to monitor and control the system, in addition to archiving the complete process history for every screen in real time. The Tox21 screening system uses a single robotic arm for plate transportation and a storage capacity of nearly 1 million compound or assay wells (about 600 1,536-well compound or assay plates) to screen smaller scale compound libraries not present on the primary screening system and the Tox21 compound library. This platform is designed to complement the primary system, and it is easily customizable, allowing for the rapid integration of additional screening technologies.
The RSC team additionally provides screening system services to intramural programs within NCATS. The Tox21 screening system contains a single robotic arm for plate transportation and a storage capacity of about 1.5 million compound wells (about 1,000 1,536-well compound plates) and more than 1.5 million assay wells (approximately 1,100 1,536-well assay plates). With a pin tool and two acoustic dispensers for compound addition, four low-volume dispensers for reagent addition, and three plate readers enabling a variety of assay detection methods, this system can rapidly screen, in triplicate, the Toxicology in the 21st Century (Tox21) library of 10,000 compounds. This year, the RSC team was also able to complete the installation of the Hamamatsu FDSS plate reader allowing the Tox21 program to expand the kinds of assays run on the platform, in particular calcium channel. The FDSS itself was modified to include a custom 1536 pintool cleaner developed by IonField; which is a first of its kind using cold plasma technology instead of DMSO and other solvents to clean, providing both better data due to minimizing carry-over effects between plates and reducing the need for caustic solvents which in the past have damaged the instrument. NCATS experts monitor and control this system with the same custom software used for the primary screening system. The RNAi screening system has two robotic arms for plate transportation with incubators for assay plate storage and plate stackers to facilitate continuous system operation. A multichannel pipettor, low volume dispensers, an aspirator for plate washing, and two different plate readers support a wide variety of detection methods for this RNAi-focused system.
RSC staff also continue to support the Stem Cell Translation Laboratory (SCTL). Various team members have provided a large amount of support with a vast array of equipment including automated tissue culture systems, flow cytometers, cell sorters, and high content imagers. Special mention should be given to the automated tissue culture platform which has established itself as a core capability for all NCATS researchers now with the demand becoming so great that another system is most likely on order. These resources require a tremendous amount of expertise to operate and maintain. In the past year, RSC has begun to act as a core facility for these technologies, enabling different biology groups to increase their capabilities. Related to the high content imagers, more high content assays were run in the past year due to the increased capabilities as compared to the previous four years combined.
The NCATS intramural program is heavily involved in COVID-19 relief efforts across the NIH. RSC is directly involved in several of these key initiatives, providing project management, scientific and technical support across several programs. Below are a list of some of the COVID-19 projects RSC has worked on:
NIH Serosurvey: A new study has begun recruiting at the National Institutes of Health in Bethesda, Maryland, to determine how many adults in the United States without a confirmed history of infection with SARS-CoV-2, the virus that causes coronavirus disease 2019 (COVID-19), have antibodies to the virus. The presence of antibodies in the blood indicates a prior infection. In this serosurvey, researchers will collect and analyze blood samples from as many as 10,000 volunteers to provide critical data for epidemiological models. The results will help illuminate the extent to which the novel coronavirus has spread undetected in the United States and provide insights into which communities and populations are most affected.
NCATS and RSC are heavily involved in this effort on two fronts. For the survey itself, RSC is creating a web-based dashboard which will directly connect to the REDCap clinical trial software system being setup at NIAID. As results from the study are entered into REDCap, RSC is creating a dashboard which will programmatically pull the most recent results and update on a daily basis for program staff to monitor progress of the study and to make sure key milestones are being met.
To analyze the samples provided by the volunteers, RSC is also assisting by providing automation and instrumentation expertise to the lab at NIBIB where the results are being processed. The diagnostic being run is an ELISA assay, with the results for patients being read out on a BMG Labtech PHERAstar plate reader. These results will also be updated to the same web-based dashboard so program staff can see this data as well. Although this initial phase is starting with 10,000 volunteers, the demand has been massive, and it is highly likely RSC will be involved with turning this currently manual process to run the ELISA assay into a fully automated process.
Compound testing in the SARS-CoV-2 CPE assay (non-GLP): This project involves sending pre-spotted assay plates prepared by the ITRB Compound Management group to run assays at the Southern Research Institute (SRI). This contract was awarded (75N95020P00144) and five rounds of assays have already been completed. This data will be made publicly available via a website that RSC will provide all hosting support for called the OpenData Portal. In addition, the next round of experiments will be to take the hits from the initial experiment and to run them in a combination dose response fashion, referred to as a matrix screen. All this plating will be performed by RSC staff and it will take place on our LabCyte Access system. As more experiments are run, more data will be made available on the COVID-19 data website and more compounds will need to be sent to SRI and potentially other BSL3 facilities.
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