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BioAssay: AID 984

Cell Viability - LYMP2-016

We have developed a 1536-well cell-based assay for quantitative high throughput screening (qHTS) against a number of cell lines to determine in vitro cytotoxicity of small molecules. This particular assay uses a human lymphoblastoid cell line (LYMP2-016) purchased from Coriell Institute for Medical Research. ..more
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 Tested Compounds
 Tested Compounds
All(1335)
 
 
Active(75)
 
 
Inactive(1186)
 
 
Inconclusive(77)
 
 
 Tested Substances
 Tested Substances
All(1408)
 
 
Active(78)
 
 
Inactive(1248)
 
 
Inconclusive(82)
 
 
 Related BioAssays
 Related BioAssays
AID: 984
Data Source: NCGC (UPGV216)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Screening Center Network
BioAssay Version:
Deposit Date: 2007-12-26
Modify Date: 2010-07-06

Data Table ( Complete ):           View Active Data    View All Data
BioActive Compounds: 75
Description:
NIH Chemical Genomics Center [NCGC]
NIH Molecular Libraries Screening Centers Network [MLSCN]
National Institutes of Environmental Health Sciences [NIEHS]
National Toxicology Program [NTP]

MLSCN Grant: None

NCGC Assay Overview:

We have developed a 1536-well cell-based assay for quantitative high throughput screening (qHTS) against a number of cell lines to determine in vitro cytotoxicity of small molecules. This particular assay uses a human lymphoblastoid cell line (LYMP2-016) purchased from Coriell Institute for Medical Research.
Protocol
NCGC Assay Protocol Summary:
The CellTiter-Glo luminescent cell viability assay (Promega) is a homogeneous method to measure the number of viable cells in culture. The end point readout of this assay is based on quantitation of intracellular ATP, an indicator of metabolic activity, using the luciferase reaction. Luciferase catalyzes the oxidation of beetle Luciferin to oxyluciferin and light in the presence of ATP. The luminescent signal is proportional to amount of ATP present.
Using the CellTiter-Glo luminescent cell viability assay, the amount of cellular ATP was measured in the lymphoblastoid cell line with complete culture medium following compound treatment for 24 hours. The assay was performed in opaque white Kalypsys 1536-well plates. In the screen, tetraoctylammonium bromide was used as positive control. Library compounds were measured for their ability to cause acute toxicity in the cell line, as reflected by a decrease in intracellular ATP levels, in a concentration-dependent manner. Data were normalized to the controls for basal activity (DMSO only) and 100% inhibition (92 uM tetraoctylammonium bromide). AC50 values were determined from concentration-response data modeled with the standard Hill equation.
qHTS protocol for CellTiter Glo lymphoblastoid cellular assay
[Step] [Parameter] [Value] [Description]
1. Reagent; 5 uL; 2500 lymphoblastoid cells/well
2. Time; 5 hr; 37 C incubation
3. Compounds; 23 nL; 0.59 nM to 92 uM
4. Controls; 23 nL; Tetraoctylammonium bromide 6.4pM to 92 uM
5. Time; 24 hr; 37 C incubation
6. Reagent; 5 uL; CellTiter Glo reagent
7. Time; 30 min; Room Temperature
8. Detection; Luminescence; Viewlux plate reader
Keywords: cell viability, cytotoxicity, lymphoblastoid cell line, luminescence, NTPHTS, NTPHTS_NCGC, NIEHS, DSSTox, MLSMR, MLSCN, NIH Roadmap, qHTS, NCGC
Comment
Compound Ranking:
1. Compounds are first classified as having full titration curves, partial modulation, partial curve (weaker actives), single point activity (at highest concentration only), or inactive. See data field "Curve Description".
2. For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds, a score range was given for each curve class type given above. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39. Fit_LogAC50 was used for determining relative score and was scaled to each curve class' score range.
Categorized Comment - additional comments and annotations
From ChEMBL:
Assay Format: Cell-based
Assay Type: Functional
Assay Cell Type: Lymphoblastoid cells
Result Definitions
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TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1PhenotypeIndicates type of activity observed: inhibitor, activator, fluorescent, cytotoxic, inactive, or inconclusive.String
2Potency*Concentration at which compound exhibits half-maximal efficacy, AC50. Extrapolated AC50s also include the highest efficacy observed and the concentration of compound at which it was observed.FloatμM
3EfficacyMaximal efficacy of compound, reported as a percentage of control. These values are estimated based on fits of the Hill equation to the dose-response curves.Float%
4Analysis CommentAnnotation/notes on a particular compound's data or its analysis.String
5Curve_DescriptionA description of dose-response curve quality. A complete curve has two observed asymptotes; a partial curve may not have attained its second asymptote at the highest concentration tested. High efficacy curves exhibit efficacy greater than 80% of control. Partial efficacies are statistically signficant, but below 80% of control.String
6Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
7Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
8Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
9Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
10Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
11Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
12Excluded_PointsWhich dose-response titration points were excluded from analysis based on outlier analysis. Each number represents whether a titration point was (1) or was not (0) excluded, for the titration series going from smallest to highest compound concentrations.String
13Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
14Activity at 0.0005903226 uM (0.000590323μM**)% Activity at given concentration.Float%
15Activity at 0.00295 uM (0.00295023μM**)% Activity at given concentration.Float%
16Activity at 0.015 uM (0.0147512μM**)% Activity at given concentration.Float%
17Activity at 0.033 uM (0.0329834μM**)% Activity at given concentration.Float%
18Activity at 0.074 uM (0.0737507μM**)% Activity at given concentration.Float%
19Activity at 0.165 uM (0.164906μM**)% Activity at given concentration.Float%
20Activity at 0.369 uM (0.368731μM**)% Activity at given concentration.Float%
21Activity at 0.824 uM (0.824482μM**)% Activity at given concentration.Float%
22Activity at 1.844 uM (1.84354μM**)% Activity at given concentration.Float%
23Activity at 4.122 uM (4.12216μM**)% Activity at given concentration.Float%
24Activity at 9.217 uM (9.21715μM**)% Activity at given concentration.Float%
25Activity at 20.61 uM (20.6095μM**)% Activity at given concentration.Float%
26Activity at 46.08 uM (46.0829μM**)% Activity at given concentration.Float%
27Activity at 92.17 uM (92.1659μM**)% Activity at given concentration.Float%
28Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.

Data Table (Concise)
Data Table ( Complete ):     View Active Data    View All Data
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