Practical Considerations of Liquid Handling Devices in Drug Discovery

Chai, Sergio C.; Goktug, Asli N.; Cui, Jimmy; Low, Jonathan; Chen, Taosheng

doi:10.5772/52546

Cited by 5 publications

(2 citation statements)

References 36 publications

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“…The presence of FBS completely prevented triciribine from enhancing FGF14:Nav1.6 complementation (10% FBS: 103.1 ± 8.9%, n = 8; 5% FBS: 97.6 ± 8.1%, n = 8; no FBS: 142.5% ± 10.7%, n = 8, p < 0.0001), and a higher concentration of FBS significantly reduced the potency of ZL181 (21.2 ± 2.8%, n = 8, p < 0.0001) compared to media with no FBS (11.2 ± 1.5%, n = 8, p < 0.0001). To circumvent this issue, as well as minimize potential variance associated with multiple liquid handling steps 35 , we attempted using cells in suspension by dispensing immediately prior to screening (cell-based homogeneous assay). Superior raw luminescence values (10446 ± 233.2 RLU, n = 8) were observed compared to adherent cells (8692 ± 78.7 RLU, n = 8, p < 0.0001, Supplementary Fig.…”

Section: Resultsmentioning

confidence: 99%

High-throughput screening against protein:protein interaction interfaces reveals anti-cancer therapeutics as potent modulators of the voltage-gated Na+ channel complex

Wadsworth

Folorunso

Nguyen

et al. 2019

Sci Rep

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Multiple voltage-gated Na+ (Nav) channelopathies can be ascribed to subtle changes in the Nav macromolecular complex. Fibroblast growth factor 14 (FGF14) is a functionally relevant component of the Nav1.6 channel complex, a causative link to spinocerebellar ataxia 27 (SCA27) and an emerging risk factor for neuropsychiatric disorders. Yet, how this protein:channel complex is regulated in the cell is still poorly understood. To search for key cellular pathways upstream of the FGF14:Nav1.6 complex, we have developed, miniaturized and optimized an in-cell assay in 384-well plates by stably reconstituting the FGF14:Nav1.6 complex using the split-luciferase complementation assay. We then conducted a high-throughput screening (HTS) of 267 FDA-approved compounds targeting known mediators of cellular signaling. Of the 65 hits initially detected, 24 were excluded based on counter-screening and cellular toxicity. Based on target analysis, potency and dose-response relationships, 5 compounds were subsequently repurchased for validation and confirmed as hits. Among those, the tyrosine kinase inhibitor lestaurtinib was highest ranked, exhibiting submicromolar inhibition of FGF14:Nav1.6 assembly. While providing evidence for a robust in-cell HTS platform that can be adapted to search for any channelopathy-associated regulatory proteins, these results lay the potential groundwork for repurposing cancer drugs for neuropsychopharmacology.

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Section: Resultsmentioning

confidence: 99%

High-throughput screening against protein:protein interaction interfaces reveals anti-cancer therapeutics as potent modulators of the voltage-gated Na+ channel complex

Wadsworth

Folorunso

Nguyen

et al. 2019

Sci Rep

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show abstract

“…Traditional liquid handlers are also limited in ways such as requiring “dead volumes” of reagents inside wells, which can increase the cost of experiments. To maintain high levels of accuracy, all automation equipment must undergo strict and frequent quality assessments (Chai et al, 2013). There is an additional investment of time required for the set-up and optimisation of new protocols, which when combined with the aforementioned issues can dissuade academic labs from purchasing such systems.…”

Section: Automationmentioning

confidence: 99%

Improving Reproducibility in Synthetic Biology

Jessop-Fabre

Sonnenschein

2019

Front. Bioeng. Biotechnol.

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Synthetic biology holds great promise to deliver transformative technologies to the world in the coming years. However, several challenges still remain to be addressed before it can deliver on its promises. One of the most important issues to address is the lack of reproducibility within research of the life sciences. This problem is beginning to be recognised by the community and solutions are being developed to tackle the problem. The recent emergence of automated facilities that are open for use by researchers (such as biofoundries and cloud labs) may be one of the ways that synthetic biologists can improve the quality and reproducibility of their work. In this perspective article, we outline these and some of the other technologies that are currently being developed which we believe may help to transform how synthetic biologists approach their research activities.

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An open-source semi-automated robotics pipeline for embryo immunohistochemistry

Fuqua

Jordan

Halavatyi

et al. 2021

Sci Rep

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A significant challenge for developmental systems biology is balancing throughput with controlled conditions that minimize experimental artifacts. Large-scale developmental screens such as unbiased mutagenesis surveys have been limited in their applicability to embryonic systems, as the technologies for quantifying precise expression patterns in whole animals has not kept pace with other sequencing-based technologies. Here, we outline an open-source semi-automated pipeline to chemically fixate, stain, and 3D-image Drosophila embryos. Central to this pipeline is a liquid handling robot, Flyspresso, which automates the steps of classical embryo fixation and staining. We provide the schematics and an overview of the technology for an engineer or someone equivalently trained to reproduce and further improve upon Flyspresso, and highlight the Drosophila embryo fixation and colorimetric or antibody staining protocols. Additionally, we provide a detailed overview and stepwise protocol for our adaptive-feedback pipeline for automated embryo imaging on confocal microscopes. We demonstrate the efficiency of this pipeline compared to classical techniques, and how it can be repurposed or scaled to other protocols and biological systems. We hope our pipeline will serve as a platform for future research, allowing a broader community of users to build, execute, and share similar experiments.

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Practical Considerations of Liquid Handling Devices in Drug Discovery

Cited by 5 publications

References 36 publications

High-throughput screening against protein:protein interaction interfaces reveals anti-cancer therapeutics as potent modulators of the voltage-gated Na+ channel complex

High-throughput screening against protein:protein interaction interfaces reveals anti-cancer therapeutics as potent modulators of the voltage-gated Na+ channel complex

Improving Reproducibility in Synthetic Biology

An open-source semi-automated robotics pipeline for embryo immunohistochemistry

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