scholarly journals Challenges in Implementing High-Density Formats for High Throughput Screening

1997 ◽  
Vol 2 (4) ◽  
pp. 12-19
Author(s):  
Don Rose ◽  
Tony Lemmo
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
High Density

High-Throughput Screening and Characterization of a High-Density Soybean Mutant Library Elucidate the Biosynthesis Pathway of Triterpenoid Saponins

2019 ◽  
Vol 60 (5) ◽  
pp. 1082-1097 ◽  
Author(s):  
Panneerselvam Krishnamurthy ◽  
Yukiko Fujisawa ◽  
Yuya Takahashi ◽  
Hanako Abe ◽  
Kentaro Yamane ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
High Density ◽  
Biosynthesis Pathway ◽  
Mutant Library ◽  
Triterpenoid Saponins

scholarly journals Identification of Novel Human High-Density Lipoprotein Receptor Up-regulators Using a Cell-Based High-Throughput Screening Assay

2007 ◽  
Vol 12 (2) ◽  
pp. 211-219 ◽  
Author(s):  
Yuan Yang ◽  
Zhongbing Zhang ◽  
Wei Jiang ◽  
Lei Gao ◽  
Guiyu Zhao ◽  
...  
Keyword(s):  
High Density Lipoprotein ◽  
Cell Line ◽  
High Throughput ◽  
Hepg2 Cells ◽  
High Throughput Screening ◽  
Density Lipoprotein ◽  
High Density ◽  
Luciferase Reporter ◽  
Stable Cell Line ◽  
Active Compounds

Scavenger receptor class B type I (SR-BI) is the high-affinity high-density lipoprotein (HDL) receptor, and CLA-1 is the human homologue of the murine SR-BI. CLA-1/SR-BI receptor has been suggested as a new preventative and/or therapeutic target for atherosclerosis due to its pivotal role in overall HDL cholesterol (HDL-C) metabolism and its antiatherogenic activity in vivo. To search for active compounds that can increase CLA-1 transcription, a novel cell-based assay was developed for application in high-throughput screening (HTS). Human hepatoma HepG2 cells were transfected with a CLA-1-promoter-luciferase reporter gene construct, and the stable transfected cell line was selected and named CLAp-LUC HepG2. With rosiglitazone as a positive control, this stable cell line was used to establish a specific CLA-1 gene expression assay in a 96-well microplate format. The evaluating parameter Z' value of 0.64 showed that this cell-based HTS assay was robust and reliable. Screening of 6000 microbial secondary metabolite crude extracts identified 8 positive strains. Between 2 identified CLA-1 up-regulators produced by actinomycete strain 04-4776, 4776B may stimulate not only the expression of CLA-1 on the transcriptional and translational levels but also the activity of CLA-1 to uptake the HDL-C in HepG2 cells. The active compounds originated from this HTS assay may be developed to drug candidates or lead compounds for new antiatherosclerosis agents.

scholarly journals High throughput screening using acoustic droplet ejection to combine protein crystals and chemical libraries on crystallization plates at high density

2015 ◽  
Vol 191 (1) ◽  
pp. 49-58 ◽  
Author(s):  
Ella Teplitsky ◽  
Karan Joshi ◽  
Daniel L. Ericson ◽  
Alexander Scalia ◽  
Jeffrey D. Mullen ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
High Density ◽  
Protein Crystals ◽  
Chemical Libraries ◽  
Droplet Ejection

scholarly journals Simple Absorbance-Based Assays for Ultra-High Throughput Screening

2001 ◽  
Vol 6 (1) ◽  
pp. 3-9 ◽  
Author(s):  
Patrick Lavery ◽  
Murray J.B. Brown ◽  
Andrew J. Pope
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
High Speed ◽  
Low Cost ◽  
High Density ◽  
Pharmaceutical Companies ◽  
Fluorescence Techniques ◽  
Enzyme Systems ◽  
Assay Performance ◽  
Standard Assay

In order to accommodate the predicted increase in screening required of successful pharmaceutical companies, miniaturized, high-speed HTS formats are necessary. Much emphasis has been placed on sensitive fluorescence techniques, but some systems, particularly enzymes interconverting small substrates, are likely to be refractory to such approaches. We show here that simple absorbance-based assays can be miniaturized to 10-,.d volumes in 1536- well microplates compatible with the requirements for ultra-high throughput screening. We demonstrate that, with low-cost hardware, assay performance is wholly predictable from the 2-fold decrease in pathlength for fully filled 1536-well plates compared to 96- and 384-well microplates. A number of enzyme systems are shown to work in this high-density format, and the inhibition parameters determined are comparable with those in standard assay formats. We also demonstrate the utility of kinetics measurements in miniaturized format with improvements in assay quality and the ability to extract detailed mechanistic information about inhibitors.

scholarly journals Development of a miniaturized 3D organoid culture platform for ultra-high-throughput screening

2020 ◽  
Vol 12 (8) ◽  
pp. 630-643 ◽  
Author(s):  
Yuhong Du ◽  
Xingnan Li ◽  
Qiankun Niu ◽  
Xiulei Mo ◽  
Min Qui ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Drug Discovery ◽  
High Throughput ◽  
High Throughput Screening ◽  
Large Scale ◽  
Genetically Engineered ◽  
High Density ◽  
Organoid Culture ◽  
3D Architecture

Abstract The recent advent of robust methods to grow human tissues as 3D organoids allows us to recapitulate the 3D architecture of tumors in an in vitro setting and offers a new orthogonal approach for drug discovery. However, organoid culturing with extracellular matrix to support 3D architecture has been challenging for high-throughput screening (HTS)-based drug discovery due to technical difficulties. Using genetically engineered human colon organoids as a model system, here we report our effort to miniaturize such 3D organoid culture with extracellular matrix support in high-density plates to enable HTS. We first established organoid culturing in a 384-well plate format and validated its application in a cell viability HTS assay by screening a 2036-compound library. We further miniaturized the 3D organoid culturing in a 1536-well ultra-HTS format and demonstrated its robust performance for large-scale primary compound screening. Our miniaturized organoid culturing method may be adapted to other types of organoids. By leveraging the power of 3D organoid culture in a high-density plate format, we provide a physiologically relevant screening platform to model tumors to accelerate organoid-based research and drug discovery.

scholarly journals Development of a Novel High-Density [3H]Hypoxanthine Scintillation Proximity Assay To Assess Plasmodium falciparum Growth

2016 ◽  
Vol 60 (10) ◽  
pp. 5949-5956 ◽  
Author(s):  
Cristina de Cózar ◽  
Iván Caballero ◽  
Gonzalo Colmenarejo ◽  
Laura M. Sanz ◽  
Emilio Álvarez-Ruiz ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
High Throughput ◽  
High Throughput Screening ◽  
Cell Activity ◽  
High Density ◽  
Whole Cell ◽  
Content Type ◽  
Scintillation Proximity Assay ◽  
Incorporation Assay ◽  
Incorporation Method

ABSTRACTThe discovery and development of new antimalarial drugs are becoming imperative because of the spread of resistance to current clinical treatments. The lack of robustly validated antimalarial targets and the difficulties with the building in of whole-cell activity in screening hits are hampering target-based approaches. However, phenotypic screens of structurally diverse molecule libraries are offering new opportunities for the identification of novel antimalarials. Several methodologies can be used to determine the whole-cellin vitropotencies of antimalarial hits. The [3H]hypoxanthine incorporation assay is considered the “gold standard” assay for measurement of the activity of antimalarial compounds against intraerythrocytic forms ofPlasmodium falciparum. However, the method has important limitations, as the assay is not amenable for high-throughput screening since it remains associated with the 96-well plate format. We have overcome this drawback by adapting the [3H]hypoxanthine incorporation method to a 384-well high-density format by coupling a homogeneous scintillation proximity assay (SPA) and thus eliminating the limiting filtration step. This SPA has been validated using a diverse set of 1,000 molecules, including both a representative set from the Tres Cantos Antimalarial Set (TCAMS) of compounds and molecules inactive against whole cells. The results were compared with those from theP. falciparumlactate dehydrogenase whole-cell assay, another method that is well established as a surrogate for parasite growth and is amenable for high-throughput screening. The results obtained demonstrate that the SPA-based [3H]hypoxanthine incorporation assay is a suitable design that is adaptable to high-throughput antimalarial drug screening and that maintains the features, robustness, and reliability of the standard filtration hypoxanthine incorporation method.

High-Density Reagent Storage Arrays for HIGH-THROUGHPUT Screening

2001 ◽  
pp. 477-479
Author(s):  
Sherri Biondi ◽  
Jeffrey Wolk ◽  
Gloria Cheng ◽  
Ravi Vijayendran ◽  
Tex Horning
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
High Density
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