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SLAS2017 Presentation by Dr. Bernhard Ellinger, Fraunhofer Institute for Molecular Biology and Applied Ecology, IME, Hamburg, Germany
Fraunhofer IME has had very good success using the Tecan Fluent® to perform fully automated screening of smaller compound batches rapidly and accurately, in parallel against multiple analytes and with multiple readouts.
Dr. Ellinger presented several different examples of screening campaigns carried out on the Fluent platform that benefited from the rapid turnaround, pipetting accuracy, lack of cross-contamination, high reproducibility, and ability to test multiple compounds, doses, and readouts in parallel that the Tecan system can achieve.The Fluent in use at Fraunhofer IME is equipped with a Carousel™ plate storage system, a Cytomat 10 incubator, and an Infinite® M1000 PRO microplate reader.
Dr. Ellinger described numerous screening applications, beginning with a thrombin protease inhibition screen to identify compounds active in stroke treatment. The results of the 4.5-hour fully automated fluorescent assay performed on 20 96-well plates and generating 7,680 data points showed good replication between compounds and controls and no plate effects.
Bernhard talks about automated compound screening in 3D cell cultures
The second example depicted liability profiling to identify novel therapeutic targets for neglected parasitic diseases. Three types of assays were performed on the Tecan Fluent: classical growth assays; kinase assays; and cytochrome P450 assays. The complex cytochrome P450 assays involved testing three enzymes in parallel and the addition of three different NADPH reagent systems. Assurance of no cross-contamination between wells was critical. The typically batch size was 19 compounds to generate dose response curves and 143 compounds for single replicates. These relatively small batch sizes were sufficient to obtain the information needed to allow the chemists to make modifications as they synthesized the next generation of compounds to feed back into this iterative cycle of screening and compound optimization. Assay results showed the Fluent system to be highly adaptable for parallel screening with rapid turnaround and high reproducibility.
Additional examples of robust, fully automated, parallel screening protocols run on the Fluent included an assay to assess HDAC inhibition, an assay capable of identifying cytostatic versus cytotoxic compounds, and the successful automation of toxicity assays in embryonic stem cells in 96-well format in which up to six compounds can be tested in multiple samples in parallel with dose-response measurements to detect cell viability. The most current project described by Dr. Ellinger is the automation of human iPS cell differentiation into tissues in hanging droplets on the Fluent.