Abstract 985: Development and validation of cell-based TFEB translocation assay in a high-content and high-throughput screening format

2019 ◽  
Author(s):  
Li Zhang ◽  
Ruili Huang ◽  
Wen Xing Ding ◽  
Menghang Xia
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Translocation Assay ◽  
Development And Validation

Development and validation of high-throughput screening assays for poly(ADP-ribose) polymerase-2 inhibitors

2014 ◽  
Vol 449 ◽  
pp. 188-194 ◽  
Author(s):  
Zhixiang Zhu ◽  
Jing Jin ◽  
Nina Xue ◽  
Xiuyun Song ◽  
Xiaoguang Chen
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Screening Assays ◽  
Development And Validation

scholarly journals Development of a High-Throughput Cytometric Screen to Identify Anti-Wolbachia Compounds: The Power of Public–Private Partnership

2019 ◽  
Vol 24 (5) ◽  
pp. 537-547
Author(s):  
Rachel H. Clare ◽  
Roger Clark ◽  
Catherine Bardelle ◽  
Paul Harper ◽  
Matthew Collier ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Neglected Tropical Diseases ◽  
Bacterial Symbiont ◽  
Tropical Diseases ◽  
Validation Process ◽  
Host Insect ◽  
Filarial Nematodes ◽  
Development And Validation ◽  
High Throughput Assay

The Anti- Wolbachia (A·WOL) consortium at the Liverpool School of Tropical Medicine (LSTM) has partnered with the Global High-Throughput Screening (HTS) Centre at AstraZeneca to create the first anthelmintic HTS for neglected tropical diseases (NTDs). The A·WOL consortium aims to identify novel macrofilaricidal drugs targeting the essential bacterial symbiont ( Wolbachia) of the filarial nematodes causing onchocerciasis and lymphatic filariasis. Working in collaboration, we have validated a robust high-throughput assay capable of identifying compounds that selectively kill Wolbachia over the host insect cell. We describe the development and validation process of this complex, phenotypic high-throughput assay and provide an overview of the primary outputs from screening the AstraZeneca library of 1.3 million compounds.

scholarly journals Development and validation of a hedgehog heparin-binding assay for high-throughput screening

MethodsX ◽  
2020 ◽  
pp. 101207
Author(s):  
David Lamson ◽  
Mark Hughes ◽  
Audrey Adcock ◽  
Ginger Smith ◽  
Kevin P. Williams
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Binding Assay ◽  
Heparin Binding ◽  
Development And Validation

scholarly journals Development of a Novel Ectonucleotidase Assay Suitable for High-Throughput Screening

2012 ◽  
Vol 17 (7) ◽  
pp. 993-998 ◽  
Author(s):  
Kris F. Sachsenmeier ◽  
Carl Hay ◽  
Erin Brand ◽  
Lori Clarke ◽  
Kim Rosenthal ◽  
...  
Keyword(s):  
Malachite Green ◽  
High Throughput ◽  
High Throughput Screening ◽  
High Performance ◽  
Adenosine Monophosphate ◽  
Screening Assay ◽  
Highly Sensitive ◽  
Development And Validation ◽  
Multiple Samples

5′-Ectonucleotidase (NT5E) catalyzes the conversion of adenosine monophosphate to adenosine and free phosphate. The role of this ectonucleotidase and its production of adenosine are linked with immune function, angiogenesis, and cancer. NT5E activity is typically assayed either by chromatographic quantification of substrates and products using high-performance liquid chromatography (HPLC) or by quantification of free phosphate using malachite green. These methods are not suitable for robust screening assays of NT5E activity. HPLC is not readily suitable for the rapid and efficient assay of multiple samples and malachite green is highly sensitive to the phosphate-containing buffers common in various media and sample buffers. Here the development and validation of a novel high-throughput ectonucleotidase screening assay are described, which makes use of a luciferase-based assay reagent, the Promega CellTiter-Glo kit, to measure the catabolism of AMP by NT5E. This multiwell plate-based assay facilitates the screening of potential ectonucleotidase antagonists and is unaffected by the presence of contaminating phosphate molecules present in screening samples.

scholarly journals Development and validation of a rapid, single-step reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) system potentially to be used for reliable and high-throughput screening of COVID-19

2020 ◽  
Author(s):  
Minghua Jiang ◽  
Weihua Pan ◽  
Amir Aratehfar ◽  
Wenjie Fang ◽  
Liyan Ling ◽  
...  
Keyword(s):  
Reverse Transcriptase ◽  
High Throughput ◽  
Predictive Value ◽  
High Throughput Screening ◽  
Lamp Assay ◽  
Single Step ◽  
Clinical Samples ◽  
Qrt Pcr ◽  
Pcr Assays ◽  
Development And Validation

AbstractObjectivesDevelopment and validation of a single-step and accurate reverse transcriptase loop-mediated isothermal amplification technique (RT-LAMP) for rapid identification of SARS COV-2 relative to commercial quantitative reverse transcriptase real-time PCR (qRT-PCR) assays to allow prompt initiation of proper medical care and containment of virus spread.MethodsPrimers showing optimal in-silico features were subjected to analytical sensitivity and specificity to assess the limit of detection (LOD) and cross-reaction with closely- and distantly-related viral species, and clinically prominent bacterial and fungal species. In order to evaluate the clinical utility, our RT-LAMP was subjected to a large number of clinical samples, including 213 negative and 47 positive patients, relative to two commercial quantitative RT-PCR assays.ResultsThe analytical specificity and sensitivity of our assay was 100% and 500 copies/ml when serial dilution performed in both water and sputum. Subjecting our RT-LAMP assay to clinical samples showed a high degree of specificity (99.5%), sensitivity (91.4%), positive predictive value (97.7%), and negative predictive value (98.1%) when used relative to qRT-PCR. Our RT-LAMP assay was two times faster than qRT-PCR and is storable at room temperature. A suspected case that later became positive tested positive using both our RT-LAMP and the two qRT-PCR assays, which shows the capability of our assay for screening purposes.ConclusionsWe present a rapid RT-LAMP assay that could extend the capacity of laboratories to process two times more clinical samples relative to qRT-PCR and potentially could be used for high-throughput screening purposes when demand is increasing at critical situations.

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