In Vitro High-Throughput Screening Assay for Modulators of Transcription

2002 ◽  
Vol 301 (1) ◽  
pp. 103-110 ◽  
Author(s):  
León F. Garcı́a-Martı́nez ◽  
Graham K. Bilter ◽  
Jun Wu ◽  
Joe O'Neill ◽  
Miguel S. Barbosa ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Screening Assay ◽  
High Throughput Screening Assay

Closing the empty anti-Babesia gibsoni drug pipeline in vitro using fluorescence-based high throughput screening assay

2020 ◽  
Vol 75 ◽  
pp. 102054 ◽  
Author(s):  
Mohamed Abdo Rizk ◽  
Shengwei Ji ◽  
Mingming Liu ◽  
Shimaa Abd El-Salam El-Sayed ◽  
Yongchang Li ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Babesia Gibsoni ◽  
Screening Assay ◽  
High Throughput Screening Assay ◽  
Drug Pipeline

scholarly journals Establishment of an In Vitro High-Throughput Screening Assay for Detecting Phospholipidosis-Inducing Potential

2005 ◽  
Vol 90 (1) ◽  
pp. 133-141 ◽  
Author(s):  
Toshihiko Kasahara ◽  
Kazuo Tomita ◽  
Hiroyuki Murano ◽  
Tsuyoshi Harada ◽  
Keisuke Tsubakimoto ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Screening Assay ◽  
High Throughput Screening Assay

scholarly journals Practical High-Throughput Method to Screen Compounds for Anthelmintic Activity against Caenorhabditis elegans

Molecules ◽  
2021 ◽  
Vol 26 (14) ◽  
pp. 4156
Author(s):  
Aya C. Taki ◽  
Joseph J. Byrne ◽  
Peter R. Boag ◽  
Abdul Jabbar ◽  
Robin B. Gasser
Keyword(s):  
Caenorhabditis Elegans ◽  
Small Molecules ◽  
High Throughput ◽  
High Throughput Screening ◽  
Anthelmintic Activity ◽  
Infrared Light ◽  
Parasitic Nematodes ◽  
Screening Assay ◽  
High Throughput Screening Assay

In the present study, we established a practical and cost-effective high throughput screening assay, which relies on the measurement of the motility of Caenorhabditis elegans by infrared light-interference. Using this assay, we screened 14,400 small molecules from the “HitFinder” library (Maybridge), achieving a hit rate of 0.3%. We identified small molecules that reproducibly inhibited the motility of C. elegans (young adults) and assessed dose relationships for a subset of compounds. Future work will critically evaluate the potential of some of these hits as candidates for subsequent optimisation or repurposing as nematocides or nematostats. This high throughput screening assay has the advantage over many previous assays in that it is cost- and time-effective to carry out and achieves a markedly higher throughput (~10,000 compounds per week); therefore, it is suited to the screening of libraries of tens to hundreds of thousands of compounds for subsequent evaluation and development. The present phenotypic whole-worm assay should be readily adaptable to a range of socioeconomically important parasitic nematodes of humans and animals, depending on their dimensions and motility characteristics in vitro, for the discovery of new anthelmintic candidates. This focus is particularly important, given the widespread problems associated with drug resistance in many parasitic worms of livestock animals globally.

scholarly journals Development and Application of a High-Throughput Screening Assay for HIV-1 Integrase Enzyme Activities

2005 ◽  
Vol 10 (6) ◽  
pp. 606-614 ◽  
Author(s):  
Sinu John ◽  
Thomas M. Fletcher ◽  
Colleen B. Jonsson
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Strand Transfer ◽  
Hiv Integrase ◽  
Chromosomal Dna ◽  
Screening Assay ◽  
Viral Dna ◽  
High Throughput Screening Assay ◽  
Hiv 1

Integrase (IN) mediates the covalent insertion of the retroviral genome into its host chromosomal DNA. This enzymatic activity can be reconstituted in vitro with short DNA oligonucleotides, which mimic a single viral DNA end, and purified IN. Herein we report a highly efficient and sensitive high-throughput screen, HIV Integrase Target SRI Assay (HITS™), for HIV-1 IN activity using 5′ biotin-labeled DNA (5′ BIO donor) and 3′ digoxygenin-labeled DNA (3′ DIG target). Following 3′ processing of the 5′ BIO donor, strand transfer proceeds with integration of the 5′ BIO donor into the 3′ DIG target. Products were captured on a streptavidin-coated microplate and the amount of DIG retained in the well was measured. The end point values, measured as absorbance, ranged from 0.9 to 1.5 for IN-mediated reactions as compared with background readings of 0.05 to 0.12. The Z factor for the assay ranged from 0.7 to 0.85. The assay was used to screen drugs in a high-throughput format, and furthermore, we adapted the assay to study mechanistic questions regarding the integration process. For example, using variations of the assay format, we showed high preference of E strand of the long terminal repeat (LTR) viral DNA as a target strand compared with its complementary A strand. The E strand is the strand processed by IN. Furthermore, we explored the reported inhibitory effect of reverse transcriptase on integration.

scholarly journals A High-Throughput Screening Assay for the Identification of Flavivirus NS5 Capping Enzyme GTP-Binding Inhibitors

2011 ◽  
Vol 16 (8) ◽  
pp. 852-861 ◽  
Author(s):  
Brian J. Geiss ◽  
Hillary J. Stahla-Beek ◽  
Amanda M. Hannah ◽  
Hamid H. Gari ◽  
Brittney R. Henderson ◽  
...  
Keyword(s):  
Drug Development ◽  
High Throughput ◽  
High Throughput Screening ◽  
Antiviral Drug ◽  
Functional Assay ◽  
Screening Assay ◽  
Gtp Binding ◽  
High Throughput Screening Assay ◽  
Capping Enzyme

There are no effective antivirals currently available for the treatment of flavivirus infection in humans. As such, the identification and characterization of novel drug target sites are critical to developing new classes of antiviral drugs. The flavivirus NS5 N-terminal capping enzyme (CE) is vital for the formation of the viral RNA cap structure, which directs viral polyprotein translation and stabilizes the 5′ end of the viral genome. The structure of the flavivirus CE has been solved, and a detailed understanding of the CE–guanosine triphosphate (GTP) and CE–RNA cap interactions is available. Because of the essential nature of the interaction for viral replication, disrupting CE–GTP binding is an attractive approach for drug development. The authors have previously developed a robust assay for monitoring CE–GTP binding in real time. They adapted this assay for high-throughput screening and performed a pilot screen of 46 323 commercially available compounds. A number of small-molecule inhibitors capable of displacing a fluorescently labeled GTP in vitro were identified, and a second functional assay was developed to identify false positives. The results presented indicate that the flavivirus CE cap-binding site is a valuable new target site for antiviral drug discovery and should be further exploited for broad-spectrum anti-flaviviral drug development.

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