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In vitro viability and cytotoxicity testing and same-well multi-parametric combinations for high throughput screening.

Niles AL, Moravec RA, Riss TL - Curr Chem Genomics (2009)

Bottom Line: In vitro cytotoxicity testing has become an integral aspect of drug discovery because it is a convenient, costeffective, and predictive means of characterizing the toxic potential of new chemical entities.The early and routine implementation of this testing is testament to its prognostic importance for humans.We also provide guidance for successful HTS implementation and discuss unique merits and detractions inherent in each method.

View Article: PubMed Central - PubMed

Affiliation: Research Department, Promega Corporation, 2800 Woods Hollow Road, Madison, WI, USA. andrew.niles@promega.com

ABSTRACT
In vitro cytotoxicity testing has become an integral aspect of drug discovery because it is a convenient, costeffective, and predictive means of characterizing the toxic potential of new chemical entities. The early and routine implementation of this testing is testament to its prognostic importance for humans. Although a plethora of assay chemistries and methods exist for 96-well formats, few are practical and sufficiently sensitive enough for application in high throughput screening (HTS). Here we briefly describe a handful of the currently most robust and validated HTS assays for accurate and efficient assessment of cytotoxic risk. We also provide guidance for successful HTS implementation and discuss unique merits and detractions inherent in each method. Lastly, we discuss the advantages of combining specific HTS compatible assays into multi-parametric, same-well formats.

No MeSH data available.


Protease Assay Sensitivity and Linearity. CytoTox-Glo™ (Promega Corporation) was added to serial dilutions of viable and non-viable Jurkat cells and luminescence measured after 30 minutes.
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Figure 4: Protease Assay Sensitivity and Linearity. CytoTox-Glo™ (Promega Corporation) was added to serial dilutions of viable and non-viable Jurkat cells and luminescence measured after 30 minutes.

Mentions: Dead cell protease detection is advantageous in HTS and secondary screening applications because of improved sensitivity, linearity with respect to different degrees of cytotoxicity, reduction in background and artifactual hits, and choice in detection format (Fig. 4). These attributes allow the technology to be applied in 1536 well formats as easily as in lower density plates [54]. The dead cell protease detection method is subject to enzymatic activity decay due to the kinetics of cytotoxicity, biomarker inhibition, and standard interferences associated with the detection molecules.


In vitro viability and cytotoxicity testing and same-well multi-parametric combinations for high throughput screening.

Niles AL, Moravec RA, Riss TL - Curr Chem Genomics (2009)

Protease Assay Sensitivity and Linearity. CytoTox-Glo™ (Promega Corporation) was added to serial dilutions of viable and non-viable Jurkat cells and luminescence measured after 30 minutes.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2802765&req=5

Figure 4: Protease Assay Sensitivity and Linearity. CytoTox-Glo™ (Promega Corporation) was added to serial dilutions of viable and non-viable Jurkat cells and luminescence measured after 30 minutes.
Mentions: Dead cell protease detection is advantageous in HTS and secondary screening applications because of improved sensitivity, linearity with respect to different degrees of cytotoxicity, reduction in background and artifactual hits, and choice in detection format (Fig. 4). These attributes allow the technology to be applied in 1536 well formats as easily as in lower density plates [54]. The dead cell protease detection method is subject to enzymatic activity decay due to the kinetics of cytotoxicity, biomarker inhibition, and standard interferences associated with the detection molecules.

Bottom Line: In vitro cytotoxicity testing has become an integral aspect of drug discovery because it is a convenient, costeffective, and predictive means of characterizing the toxic potential of new chemical entities.The early and routine implementation of this testing is testament to its prognostic importance for humans.We also provide guidance for successful HTS implementation and discuss unique merits and detractions inherent in each method.

View Article: PubMed Central - PubMed

Affiliation: Research Department, Promega Corporation, 2800 Woods Hollow Road, Madison, WI, USA. andrew.niles@promega.com

ABSTRACT
In vitro cytotoxicity testing has become an integral aspect of drug discovery because it is a convenient, costeffective, and predictive means of characterizing the toxic potential of new chemical entities. The early and routine implementation of this testing is testament to its prognostic importance for humans. Although a plethora of assay chemistries and methods exist for 96-well formats, few are practical and sufficiently sensitive enough for application in high throughput screening (HTS). Here we briefly describe a handful of the currently most robust and validated HTS assays for accurate and efficient assessment of cytotoxic risk. We also provide guidance for successful HTS implementation and discuss unique merits and detractions inherent in each method. Lastly, we discuss the advantages of combining specific HTS compatible assays into multi-parametric, same-well formats.

No MeSH data available.