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BioAssay: AID 1815

qHTS for differential inhibitors of proliferation of Plasmodium falciparum line 7G8

Plasmodium falciparum is a virulent malarial parasite that kills 1-2 million people worldwide each year. Transmitted by mosquitoes, the sporozoite form of the parasite enters the blood stream to infect hepatocytes, where they become merozoites that infect red blood cells (erythrocytes). Within erythrocytes, merozoites begin a 72-hour cycle of growth and division, after which the parasite is more ..
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 Tested Compounds
 Tested Compounds
All(1272)
 
 
Active(441)
 
 
Inactive(558)
 
 
Inconclusive(276)
 
 
 Tested Substances
 Tested Substances
All(1279)
 
 
Active(443)
 
 
Inactive(560)
 
 
Inconclusive(276)
 
 
 Related BioAssays
 Related BioAssays
AID: 1815
Data Source: NCGC (SYBR7G8)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2009-06-16

Data Table ( Complete ):           View Active Data    View All Data
BioActive Compounds: 441
Related Experiments
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AIDNameTypeComment
1828qHTS for Inhibitors of Plasmodium falciparum proliferation: SummarySummarydepositor-specified cross reference
1876qHTS for differential inhibitors of proliferation of Plasmodium falciparum line 3D7Confirmatorydepositor-specified cross reference
1877qHTS for differential inhibitors of proliferation of Plasmodium falciparum line D10Confirmatorydepositor-specified cross reference
1882qHTS for differential inhibitors of proliferation of Plasmodium falciparum line Dd2Confirmatorydepositor-specified cross reference
1883qHTS for differential inhibitors of proliferation of Plasmodium falciparum line W2Confirmatorydepositor-specified cross reference
1886qHTS for differential inhibitors of proliferation of Plasmodium falciparum line HB3Confirmatorydepositor-specified cross reference
1816qHTS for differential inhibitors of proliferation of Plasmodium falciparum line GB4Confirmatorysame project related to Summary assay
504314qHTS for Inhibitors of Plasmodium falciparum (DD2) proliferationConfirmatorysame project related to Summary assay
504315qHTS for Inhibitors of Plasmodium falciparum (GB4) proliferationConfirmatorysame project related to Summary assay
504316qHTS for Inhibitors of Plasmodium falciparum (7G8) proliferationConfirmatorysame project related to Summary assay
504318qHTS for Inhibitors of Plasmodium falciparum (HB3) proliferationConfirmatorysame project related to Summary assay
504320qHTS for Inhibitors of Plasmodium falciparum (CP250) proliferationConfirmatorysame project related to Summary assay
Description:
NIH Molecular Libraries Probe Production Network [MLPCN]
NIH National Institute of Allergy and Infectious Diseases [NIAID]
NIH Chemical Genomics Center [NCGC]

MLPCN Grant: None
Assay Provider: Xinzhuan Su, NIAID

NCGC Assay Overview:

Plasmodium falciparum is a virulent malarial parasite that kills 1-2 million people worldwide each year. Transmitted by mosquitoes, the sporozoite form of the parasite enters the blood stream to infect hepatocytes, where they become merozoites that infect red blood cells (erythrocytes). Within erythrocytes, merozoites begin a 72-hour cycle of growth and division, after which the parasite is released to infect new red blood cells. Gametocytes, which form within erythrocytes as well, circulate in the bloodstream and are ingested by the mosquito, thereby completing the life cycle. To profile malarial isolates for differential chemical responses and to identify inhibitors of malarial growth, an assay that measures P. falciparum growth and replication within erythrocytes was employed. This assay uses a DNA binding fluorescent dye (SYBR Green) to detect parasite DNA following cell lysis. Because mature erythrocytes lack DNA, only Plasmodium-derived DNA is detected. The P. falciparum line 7G8, derived from a malarial isolate from Brazil, was used.
Protocol
NCGC Assay Protocol Summary:
The SYBR Green viability assay was adapted from a method described previously (Plouffe et al., 2008). For screening 3 ul culture medium was dispensed into 1536 well black clear bottom plates (Aurora Biotechnologies) using a Multidrop Combi (Thermo Fisher Scientific Inc.), 23 nL compounds in DMSO were added by a pin tool (Kalypsys), and 5 ul of erythrocytes infected with P. falciparum line 7G8 (0.3% parasitemia, 2.5% hematocrit final concentration) were added. The plates were incubated at 37 C in a humidified incubator in 5% CO2 for 72 h, and 2 ul lysis buffer (20 mM Tris HCl, 10 mM EDTA, 0.16% saponin, 1.6% triton X, 10X SYBR Green I supplied as 10,000X final concentration by Invitrogen) was added to each well. The plates were mixed for 25 sec with gentle shaking and incubated overnight at room temperature in the dark. The following morning, fluorescence intensity at 485(14) nm excitation and 535(25) nm emission wavelengths was measured on an EnVision (Perkin Elmer) plate reader. Plate reads were normalized relative to the control inhibitor (0.29 uM artemisinin) and vehicle (DMSO) wells present on each plate and then corrected by an algorithm using vehicle-only control plates at the beginning and end of the compound plate stack.
References
Plouffe D, Brinker A, McNamara C, Henson K, Kato N, Kuhen K, Nagle A, Adrian F, Matzen JT, Anderson P, Nam TG, Gray NS, Chatterjee A, Janes J, Yan SF, Trager R, Caldwell JS, Schultz PG, Zhou Y, Winzeler EA. In silico activity profiling reveals the mechanism of action of antimalarials discovered in a high-throughput screen. Proc Natl Acad Sci U S A. 2008, 105(26), pp 9059-64.
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". For this assay, apparent inhibitors are ranked higher than compounds that showed apparent activation.
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 Type: Functional
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 significant, 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 0.0003681034 uM (0.000368103μM**)% Activity at given concentration.Float%
15Activity at 0.00184 uM (0.00183966μM**)% Activity at given concentration.Float%
16Activity at 0.00920 uM (0.00919828μM**)% Activity at given concentration.Float%
17Activity at 0.046 uM (0.045977μM**)% Activity at given concentration.Float%
18Activity at 0.230 uM (0.229914μM**)% Activity at given concentration.Float%
19Activity at 1.149 uM (1.14943μM**)% Activity at given concentration.Float%
20Activity at 5.747 uM (5.74713μM**)% Activity at given concentration.Float%
21Activity at 28.74 uM (28.7356μM**)% Activity at given concentration.Float%
22Compound 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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