Bioluminescent, Nonlytic, Real-Time Cell Viability Assay and Use in Inhibitor Screening

Sarah J Duellman; Wenhui Zhou; Poncho Meisenheimer; Gediminas Vidugiris; James J Cali; Prson Gautam; Krister Wennerberg; Jolanta Vidugiriene

doi:10.1089/adt.2015.669

Bioluminescent, Nonlytic, Real-Time Cell Viability Assay and Use in Inhibitor Screening

Sarah J Duellman, Wenhui Zhou, Poncho Meisenheimer, Gediminas Vidugiris, James J Cali, Prson Gautam, Krister Wennerberg, Jolanta Vidugiriene

26 Citationer (Scopus)

Abstract

Real-time continuous monitoring of cellular processes offers distinct advantages over traditional endpoint assays. A comprehensive representation of the changes occurring in live cells over the entire length of an experiment provides information about the biological status of the cell and informs decisions about the timing of treatments or the use of other functional endpoint assays. We describe a homogeneous, nonlytic, bioluminescent assay that measures cell viability in real time. This time-dependent measurement allowed us to monitor cell health for 72 h from the same test samples, distinguish differential cell growth, and investigate drug mechanism of action by analyzing time- and dose-dependent drug effects. The real-time measurements also allowed us to detect cell death immediately (>75% signal decrease within 15 min of digitonin addition), analyze drug potency versus efficacy, and identify cytostatic versus toxic drug effects. We screened an oncology compound library (Z' = 0.7) and identified compounds with varying activity at different time points (1.6% of the library showed activity within 3 h, whereas 35.4% showed a response by 47 h). The assay compared well with orthogonal endpoint cell viability assays and additionally provided data at multiple time points and the opportunity to multiplex assays on the same cells. To test the advantage of time-dependent measurements to direct optimal timing of downstream applications, we used the real-time cell viability assay to determine the ideal time to measure caspase activity by monitoring the onset of cell death and multiplexing a luminescent caspase activation assay on the same test samples.

Originalsprog	Engelsk
Tidsskrift	ASSAY and Drug Development Technologies
Vol/bind	13
Udgave nummer	8
Sider (fra-til)	456-65
Antal sider	10
ISSN	1540-658X
DOI	https://doi.org/10.1089/adt.2015.669
Status	Udgivet - 1 okt. 2015
Udgivet eksternt	Ja

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10.1089/adt.2015.669

adt.2015.669Forlagets udgivne version, 613 KB

Citationsformater

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abstract = "Real-time continuous monitoring of cellular processes offers distinct advantages over traditional endpoint assays. A comprehensive representation of the changes occurring in live cells over the entire length of an experiment provides information about the biological status of the cell and informs decisions about the timing of treatments or the use of other functional endpoint assays. We describe a homogeneous, nonlytic, bioluminescent assay that measures cell viability in real time. This time-dependent measurement allowed us to monitor cell health for 72 h from the same test samples, distinguish differential cell growth, and investigate drug mechanism of action by analyzing time- and dose-dependent drug effects. The real-time measurements also allowed us to detect cell death immediately (>75% signal decrease within 15 min of digitonin addition), analyze drug potency versus efficacy, and identify cytostatic versus toxic drug effects. We screened an oncology compound library (Z' = 0.7) and identified compounds with varying activity at different time points (1.6% of the library showed activity within 3 h, whereas 35.4% showed a response by 47 h). The assay compared well with orthogonal endpoint cell viability assays and additionally provided data at multiple time points and the opportunity to multiplex assays on the same cells. To test the advantage of time-dependent measurements to direct optimal timing of downstream applications, we used the real-time cell viability assay to determine the ideal time to measure caspase activity by monitoring the onset of cell death and multiplexing a luminescent caspase activation assay on the same test samples.",

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AU - Cali, James J

AU - Gautam, Prson

AU - Wennerberg, Krister

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AB - Real-time continuous monitoring of cellular processes offers distinct advantages over traditional endpoint assays. A comprehensive representation of the changes occurring in live cells over the entire length of an experiment provides information about the biological status of the cell and informs decisions about the timing of treatments or the use of other functional endpoint assays. We describe a homogeneous, nonlytic, bioluminescent assay that measures cell viability in real time. This time-dependent measurement allowed us to monitor cell health for 72 h from the same test samples, distinguish differential cell growth, and investigate drug mechanism of action by analyzing time- and dose-dependent drug effects. The real-time measurements also allowed us to detect cell death immediately (>75% signal decrease within 15 min of digitonin addition), analyze drug potency versus efficacy, and identify cytostatic versus toxic drug effects. We screened an oncology compound library (Z' = 0.7) and identified compounds with varying activity at different time points (1.6% of the library showed activity within 3 h, whereas 35.4% showed a response by 47 h). The assay compared well with orthogonal endpoint cell viability assays and additionally provided data at multiple time points and the opportunity to multiplex assays on the same cells. To test the advantage of time-dependent measurements to direct optimal timing of downstream applications, we used the real-time cell viability assay to determine the ideal time to measure caspase activity by monitoring the onset of cell death and multiplexing a luminescent caspase activation assay on the same test samples.

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KW - Apoptosis/drug effects

KW - Cell Survival/drug effects

KW - Computer Systems

KW - Dose-Response Relationship, Drug

KW - Drug Evaluation, Preclinical/methods

KW - Humans

KW - K562 Cells

KW - Luminescent Measurements/methods

KW - Small Molecule Libraries/analysis

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JO - Assay and Drug Development Technologies

JF - Assay and Drug Development Technologies

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ER -

Bioluminescent, Nonlytic, Real-Time Cell Viability Assay and Use in Inhibitor Screening

Abstract

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