Alternative Statistical Parameter for High-Throughput Screening Assay Quality Assessment

Sui, Yunxia; Wu, Zhijin

doi:10.1177/1087057106296498

Cited by 113 publications

(110 citation statements)

References 6 publications

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“…The Z-factor for the parameter tube total length is 0.68 and thus, meets the standards for high-throughput screening (30). The value of the Z-factor is dependent on the number of EC seeded in each well.…”

Section: Image Analysis Parameters For a High-throughput Screenmentioning

confidence: 72%

A novel imaging‐based high‐throughput screening approach to anti‐angiogenic drug discovery

et al. 2009

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The successful progression to the clinic of angiogenesis inhibitors for cancer treatment has spurred interest in developing new classes of anti-angiogenic compounds. The resulting surge in available candidate therapeutics highlights the need for robust, highthroughput angiogenesis screening systems that adequately capture the complexity of new vessel formation while providing quantitative evaluation of the potency of these agents. Available in vitro angiogenesis assays are either cumbersome, impeding adaptation to high-throughput screening formats, or inadequately model the complex multistep process of new vessel formation. We therefore developed an organotypic endothelial-mural cell co-culture assay system that reflects several facets of angiogenesis while remaining compatible with high-throughput/high-content image screening. Co-culture of primary human endothelial cells (EC) and vascular smooth muscle cells (vSMC) results in assembly of a network of tubular endothelial structures enveloped with vascular basement membrane proteins, thus, comprising the three main components of blood vessels. Initially, EC are dependent on vSMC-derived VEGF and sensitive to clinical anti-angiogenic therapeutics. A subsequent phenotypic VEGFswitch renders EC networks resistant to anti-VEGF therapeutics, demarcating a mature vascular phenotype. Conversely, mature EC networks remain sensitive to vascular disrupting agents. Therefore, candidate anti-angiogenic compounds can be interrogated for their relative potency on immature and mature networks and classified as either vascular normalizing or vascular disrupting agents. Here, we demonstrate that the EC-vSMC co-culture assay represents a robust high-content imaging high-throughput screening system for identification of novel anti-angiogenic agents. A pilot highthroughput screening campaign was used to define informative imaging parameters and develop a follow-up dose-response scheme for hit characterization. High-throughput screening using the EC-vSMC co-culture assay establishes a new platform to screen for novel anti-angiogenic compounds for cancer therapy. ' 2009 International Society for Advancement of Cytometry Key termshigh-throughput screening; endothelial/mural cell co-culture; angiogenesis INHIBITING angiogenesis is a validated therapeutic modality for cancer (1). Hence, new agents that potently inhibit pathological angiogenesis are a critical component of future combination cancer therapies (2). The identification of candidate therapeutics relies on current in vitro angiogenesis assays that measure effects on endothelial cell migration, proliferation, apoptosis and tube formation, and in vivo models such as the chick chorioallantoic membrane assay, corneal neovascularization assay, and Matrigel plug assays (3-5). Current in vitro angiogenesis assays are generally focused on specific behaviors of monocultured endothelial cells (EC) (e.g., migration, proliferation) and fail to model heterotypic perivascular interactions, whereas in vivo systems are not compatible ...

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Section: Image Analysis Parameters For a High-throughput Screenmentioning

confidence: 72%

A novel imaging‐based high‐throughput screening approach to anti‐angiogenic drug discovery

et al. 2009

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“…19 A Z′ factor > 0.5 is often interpreted as an indicator of acceptable assay quality. 20 For any assay, if CV is > 0.17 (17%), the Z′ factor will not reach 0.5. 20 Thus, an assay will become less amenable to HTS.…”

Section: Introductionmentioning

confidence: 99%

“…20 For any assay, if CV is > 0.17 (17%), the Z′ factor will not reach 0.5. 20 Thus, an assay will become less amenable to HTS. A CV value closer to 0 will reflect a more reproducible assay for application to HTS.…”

Section: Introductionmentioning

confidence: 99%

Development of an Alamar Blue™ Viability Assay in 384-Well Format for High Throughput Whole Cell Screening of Trypanosoma brucei brucei Bloodstream Form Strain 427

Sykes¹,

Avery²

2009

The American Journal of Tropical Medicine and Hygiene

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Abstract. There is an urgent need for new compounds for the drug development pipeline for treatment of patients with African sleeping sickness. One approach for identifying such compounds is by high throughput screening (HTS) of compound collections. For time and cost considerations, there is a need for the development of an assay that uses at least 384-well formats. To our knowledge, there are currently no viability assays for whole cell screening of trypanosomes in the 384-well plate format. We have developed and optimized an Alamar Blue viability assay in a 384-well format for Trypanosoma brucei brucei bloodstream form strain 427 (BS427). The assay had a Z′ > 0.5 and tolerated a final dimethylsulfoxide concentration of 0.42%. Drug sensitivity was compared with those reported from previously developed 96-well methods and was found to be comparable. The sensitivity and cost benefit of the Alamar Blue assay make it an excellent candidate for HTS application.

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“…26,34,35 SSMD has recently been proposed for assessing data quality in RNAi HTS assays. 28 Compared to Z factor, SSMD is more favorable in terms of both probabilistic interpretation and statistical estimation and inference.…”

Section: Figmentioning

confidence: 99%

Integrating Experimental and Analytic Approaches to Improve Data Quality in Genome-wide RNAi Screens

Zhang¹,

Espeseth²,

Johnson³

et al. 2008

SLAS Discovery

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Alternative Statistical Parameter for High-Throughput Screening Assay Quality Assessment

Cited by 113 publications

References 6 publications

A novel imaging‐based high‐throughput screening approach to anti‐angiogenic drug discovery

A novel imaging‐based high‐throughput screening approach to anti‐angiogenic drug discovery

Development of an Alamar Blue™ Viability Assay in 384-Well Format for High Throughput Whole Cell Screening of Trypanosoma brucei brucei Bloodstream Form Strain 427

Integrating Experimental and Analytic Approaches to Improve Data Quality in Genome-wide RNAi Screens

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