scholarly journals High-throughput screening for developability during early-stage antibody discovery using self-interaction nanoparticle spectroscopy

mAbs ◽  
2013 ◽  
Vol 6 (2) ◽  
pp. 483-492 ◽  
Author(s):  
Yuqi Liu ◽  
Isabelle Caffry ◽  
Jiemin Wu ◽  
Steven B Geng ◽  
Tushar Jain ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Early Stage ◽  
Antibody Discovery

scholarly journals Cytotoxic Profiling of Annotated and Diverse Chemical Libraries Using Quantitative High-Throughput Screening

2019 ◽  
Vol 25 (1) ◽  
pp. 9-20 ◽  
Author(s):  
Olivia W. Lee ◽  
Shelley Austin ◽  
Madison Gamma ◽  
Dorian M. Cheff ◽  
Tobie D. Lee ◽  
...  
Keyword(s):  
Cell Lines ◽  
Cancer Cell ◽  
High Throughput ◽  
High Throughput Screening ◽  
Drug Targets ◽  
Large Scale ◽  
Early Stage ◽  
Cancer Cell Line ◽  
Hek 293 ◽  
Cytotoxic Compounds

Cell-based phenotypic screening is a commonly used approach to discover biological pathways, novel drug targets, chemical probes, and high-quality hit-to-lead molecules. Many hits identified from high-throughput screening campaigns are ruled out through a series of follow-up potency, selectivity/specificity, and cytotoxicity assays. Prioritization of molecules with little or no cytotoxicity for downstream evaluation can influence the future direction of projects, so cytotoxicity profiling of screening libraries at an early stage is essential for increasing the likelihood of candidate success. In this study, we assessed the cell-based cytotoxicity of nearly 10,000 compounds in the National Institutes of Health, National Center for Advancing Translational Sciences annotated libraries and more than 100,000 compounds in a diversity library against four normal cell lines (HEK 293, NIH 3T3, CRL-7250, and HaCat) and one cancer cell line (KB 3-1, a HeLa subline). This large-scale library profiling was analyzed for overall screening outcomes, hit rates, pan-activity, and selectivity. For the annotated library, we also examined the primary targets and mechanistic pathways regularly associated with cell death. To our knowledge, this is the first study to use high-throughput screening to profile a large screening collection (>100,000 compounds) for cytotoxicity in both normal and cancer cell lines. The results generated here constitute a valuable resource for the scientific community and provide insight into the extent of cytotoxic compounds in screening libraries, allowing for the identification and avoidance of compounds with cytotoxicity during high-throughput screening campaigns.

scholarly journals Cytotoxic Profiling of Annotated and Diverse Chemical Libraries Using Quantitative High-Throughput Screening

2018 ◽  
Author(s):  
Olivia W. Lee ◽  
Shelley Austin ◽  
Madison Gamma ◽  
Dorian M. Cheff ◽  
Tobie D. Lee ◽  
...  
Keyword(s):  
Cell Lines ◽  
Cancer Cell ◽  
High Throughput ◽  
High Throughput Screening ◽  
Drug Targets ◽  
Large Scale ◽  
Early Stage ◽  
Cancer Cell Line ◽  
Hek 293 ◽  
Cytotoxic Compounds

AbstractCell-based phenotypic screening is a commonly used approach to discover biological pathways, novel drug targets, chemical probes and high-quality hit-to-lead molecules. Many hits identified from high-throughput screening campaigns are ruled out through a series of follow-up potency, selectivity/specificity, and cytotoxicity assays. Prioritization of molecules with little or no cytotoxicity for downstream evaluation can influence the future direction of projects, so cytotoxicity profiling of screening libraries at an early stage is essential for increasing the likelihood of candidate success. In this study, we assessed cell-based cytotoxicity of nearly 10,000 compounds in NCATS annotated libraries, and over 100,000 compounds in a diversity library, against four ‘normal’ cell lines (HEK 293, NIH 3T3, CRL-7250 and HaCat) and one cancer cell line (KB 3-1, a HeLa subline). This large-scale library profiling was analyzed for overall screening outcomes, hit rates, pan-activity and selectivity. For the annotated library, we also examined the primary targets and mechanistic pathways regularly associated with cell death. To our knowledge, this is the first study to use high-throughput screening to profile a large screening collection (>100,000 compounds) for cytotoxicity in both normal and cancer cell lines. The results generated here constitutes a valuable resource for the scientific community and provides insight on the extent of cytotoxic compounds in screening libraries, identifying and avoiding compounds with cytotoxicity during high-throughput screening campaigns.

scholarly journals Development of a Fluorescence-Based, Ultra High-Throughput Screening Platform for Nanoliter-Scale Cytochrome P450 Microarrays

2009 ◽  
Vol 14 (6) ◽  
pp. 668-678 ◽  
Author(s):  
Sumitra M. Sukumaran ◽  
Benjamin Potsaid ◽  
Moo-Yeal Lee ◽  
Douglas S. Clark ◽  
Jonathan S. Dordick
Keyword(s):  
Enzyme Activity ◽  
Cytochrome P450 ◽  
High Throughput ◽  
High Throughput Screening ◽  
Imaging System ◽  
Early Stage ◽  
Wide Field ◽  
Cytochrome P450 Enzyme ◽  
Assay Sensitivity ◽  
Screening Platform

Cytochrome P450 enzyme (CYP450s) assays are critical enzymes in early-stage lead discovery and optimization in drug development. Currently available fluorescence-based reaction assays provide a rapid and reliable method for monitoring CYP450 enzyme activity but are confined to medium-throughput well-plate systems. The authors present a high-throughput, integrated screening platform for CYP450 assays combining enzyme encapsulation techniques, microarraying methods, and wide-field imaging. Alginate-containing microarrays consisting of up to 1134 CYP450 reaction elements were fabricated on functionalized glass slides (reaction volumes 20 to 80 nL, total enzyme content in pg) and imaged to yield endpoint activity, stability, and kinetic data. A charge-coupled device imager acquired quantitative, high-resolution images of a 20 × 20 mm area/snapshot using custom-built wide-field optics with telecentric lenses and easily interchangeable filter sets. The imaging system offered a broad dynamic intensity range (linear over 3 orders of magnitude) and sensitivity down to fluorochrome quantities of <5 fmols, with read accuracy similar to a laser scanner or a fluorescence plate reader but with higher throughput. Rapid image acquisition enabled analysis of CYP450 kinetics. Fluorogenic assays with CYP3A4, CYP2C9, and CYP2D6 on the alginate microarrays exhibited Z′ factors ranging from 0.75 to 0.85, sensitive detection of inhibitory compounds, and reactivity comparable to that in solution, thereby demonstrating the reliability and accuracy of the microarray platform. This system enables for the first time a significant miniaturization of CYP enzyme assays with significant conservation of assay reagents, greatly increased throughput, and no apparent loss of enzyme activity or assay sensitivity. ( Journal of Biomolecular Screening 2009:668-678)

scholarly journals High throughput screening of Toxoplasma gondii clones by PCR after limitation cloning

2013 ◽  
Author(s):  
Ting-Kai Liu
Keyword(s):  
Toxoplasma Gondii ◽  
High Throughput ◽  
High Throughput Screening ◽  
Early Stage ◽  
Gene Knockout ◽  
Obligate Intracellular ◽  
Intracellular Parasites ◽  
Serological Studies ◽  
Gene Defects ◽  
Global Population

Toxoplasma gondii is a kind of obligate intracellular parasites that are capable of infecting virtually all warm-blooded animals. It is one of the most common parasites in human. Serological studies estimated that up to a third of the global population has been chronically infected with the parsites. Toxoplasma gondii is also used as a model orgnisim of Apicomplexans that includes Plasmodium – the parasites that cause malaria. Gene knockout is a very important way to study gene function in all organisms. When it comes to Toxoplasma, it’s very difficult as this haploid parasite has strong adaptability to circumvent the condition of gene defects by gene duplication. High throughput screening at very early stage of transfection is very important for generating a true knockout of this parasite. For some genes that are required forToxoplasma gondii, the high throughput screening is necessary. Different labs use different protocols. After reading and practicing protocols from different papers or different labs, here I established an efficient pipeline for this purpose.

scholarly journals High throughput screening of Toxoplasma gondii clones by PCR after limitation cloning

2013 ◽  
Author(s):  
Ting-Kai Liu
Keyword(s):  
Toxoplasma Gondii ◽  
High Throughput ◽  
High Throughput Screening ◽  
Early Stage ◽  
Gene Knockout ◽  
Obligate Intracellular ◽  
Intracellular Parasites ◽  
Serological Studies ◽  
Gene Defects ◽  
Global Population

Toxoplasma gondii is a kind of obligate intracellular parasites that are capable of infecting virtually all warm-blooded animals. It is one of the most common parasites in human. Serological studies estimated that up to a third of the global population has been chronically infected with the parsites. Toxoplasma gondii is also used as a model orgnisim of Apicomplexans that includes Plasmodium – the parasites that cause malaria. Gene knockout is a very important way to study gene function in all organisms. When it comes to Toxoplasma, it’s very difficult as this haploid parasite has strong adaptability to circumvent the condition of gene defects by gene duplication. High throughput screening at very early stage of transfection is very important for generating a true knockout of this parasite. For some genes that are required forToxoplasma gondii, the high throughput screening is necessary. Different labs use different protocols. After reading and practicing protocols from different papers or different labs, here I established an efficient pipeline for this purpose.

Utility of High Throughput Screening Techniques to Predict Stability of Monoclonal Antibody Formulations During Early Stage Development

2017 ◽  
Vol 106 (8) ◽  
pp. 1971-1977 ◽  
Author(s):  
Deborah S. Goldberg ◽  
Rachael A. Lewus ◽  
Reza Esfandiary ◽  
David C. Farkas ◽  
Neil Mody ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
High Throughput ◽  
High Throughput Screening ◽  
Early Stage ◽  
Screening Techniques ◽  
Stage Development

scholarly journals Zebrafish Model for Screening Antiatherosclerosis Drugs

2021 ◽  
Vol 2021 ◽  
pp. 1-13
Author(s):  
Jichun Han ◽  
Rui Zhang ◽  
Xiaofeng Zhang ◽  
Jing Dong ◽  
Minghan Chen ◽  
...  
Keyword(s):  
Lipid Metabolism ◽  
High Throughput ◽  
High Throughput Screening ◽  
Early Stage ◽  
Vascular Inflammation ◽  
Fibrous Tissue ◽  
High Cholesterol Diet ◽  
Zebrafish Model ◽  
Lipid Metabolism Disorder ◽  
Lipid Metabolism Disorders

This study is aimed at establishing a zebrafish model of AS, which can be applied for high-throughput screening anti-AS drugs. A zebrafish AS model was induced by high cholesterol diet (HCD) and lipopolysaccharide (LPS). In the early stage of modeling, HCD induced zebrafish to show some early symptoms similar to human AS, mainly cholesterol accumulation, vascular inflammation, lipid metabolism disorder, and oxidative stress. In addition to lipid metabolism disorders, LPS also induced the same symptoms. And when HCD and LPS exist at the same time, these AS symptoms in zebrafish become more severe. When the modeling time reached 45 days, HCD and LPS induce the formation of plaques in zebrafish blood vessels, and these plaques contain fibrous tissue and lipids, which are similar to human AS plaques. We also evaluated the efficacy of some anti-AS drugs (atorvastatin, aspirin, and vitamin C) through these zebrafish AS models. The results found that atorvastatin can significantly reduce the symptoms of AS induced by HCD and LPS, and aspirin and vitamins can significantly reduce the symptoms of AS induced by LPS. It is feasible to use zebrafish to establish an AS model, and the zebrafish AS model can be used for high-throughput screening of anti-AS drugs.

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