Abstract
The emergence and spread of multidrug-resistant Plasmodium falciparum and recent detection of potential artemisinin-resistant strains in Southeast Asia highlight the importance of developing novel antimalarial therapies. Using a previously generated stable transgenic P. falciparum line with high-level firefly luciferase expression, we report the adaptation, miniaturization, optimization, and validation of a high-throughput screening assay in 384-well plates. Assay conditions, including the percentage of parasitemia and hematocrit, were optimized. Parameters of assay robustness, including Z′-value, coefficient variation (CV), and signal-to-background (S/B) ratio, were determined. The LOPAC 1280 small-compound library was used to validate this assay. Our results demonstrated that this assay is robust and reliable, with an average Z′-value of >0.7 and CV of <10%. Moreover, this assay showed a very low background, with the S/B ratio up to 71. Further, identified hits were selected and confirmed using a SYBR Green I-based confirmatory assay. It is evident that this assay is suitable for large-scale screening of chemical libraries for antimalarial drug discovery.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 61-68 |
| Number of pages | 8 |
| Journal | Assay and Drug Development Technologies |
| Volume | 10 |
| Issue number | 1 |
| DOIs | |
| State | Published - Feb 1 2012 |
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All Science Journal Classification (ASJC) codes
- Molecular Medicine
- Drug Discovery
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Validating a firefly luciferase-based high-throughput screening assay for antimalarial drug discovery. / Che, Pulin; Cui, Long; Kutsch, Olaf; Cui, Liwang; Li, Qianjun.
In: Assay and Drug Development Technologies, Vol. 10, No. 1, 01.02.2012, p. 61-68.Research output: Contribution to journal › Article
TY - JOUR
T1 - Validating a firefly luciferase-based high-throughput screening assay for antimalarial drug discovery
AU - Che, Pulin
AU - Cui, Long
AU - Kutsch, Olaf
AU - Cui, Liwang
AU - Li, Qianjun
PY - 2012/2/1
Y1 - 2012/2/1
N2 - The emergence and spread of multidrug-resistant Plasmodium falciparum and recent detection of potential artemisinin-resistant strains in Southeast Asia highlight the importance of developing novel antimalarial therapies. Using a previously generated stable transgenic P. falciparum line with high-level firefly luciferase expression, we report the adaptation, miniaturization, optimization, and validation of a high-throughput screening assay in 384-well plates. Assay conditions, including the percentage of parasitemia and hematocrit, were optimized. Parameters of assay robustness, including Z′-value, coefficient variation (CV), and signal-to-background (S/B) ratio, were determined. The LOPAC 1280 small-compound library was used to validate this assay. Our results demonstrated that this assay is robust and reliable, with an average Z′-value of >0.7 and CV of <10%. Moreover, this assay showed a very low background, with the S/B ratio up to 71. Further, identified hits were selected and confirmed using a SYBR Green I-based confirmatory assay. It is evident that this assay is suitable for large-scale screening of chemical libraries for antimalarial drug discovery.
AB - The emergence and spread of multidrug-resistant Plasmodium falciparum and recent detection of potential artemisinin-resistant strains in Southeast Asia highlight the importance of developing novel antimalarial therapies. Using a previously generated stable transgenic P. falciparum line with high-level firefly luciferase expression, we report the adaptation, miniaturization, optimization, and validation of a high-throughput screening assay in 384-well plates. Assay conditions, including the percentage of parasitemia and hematocrit, were optimized. Parameters of assay robustness, including Z′-value, coefficient variation (CV), and signal-to-background (S/B) ratio, were determined. The LOPAC 1280 small-compound library was used to validate this assay. Our results demonstrated that this assay is robust and reliable, with an average Z′-value of >0.7 and CV of <10%. Moreover, this assay showed a very low background, with the S/B ratio up to 71. Further, identified hits were selected and confirmed using a SYBR Green I-based confirmatory assay. It is evident that this assay is suitable for large-scale screening of chemical libraries for antimalarial drug discovery.
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UR - http://www.scopus.com/inward/citedby.url?scp=84863233246&partnerID=8YFLogxK
U2 - 10.1089/adt.2011.0378
DO - 10.1089/adt.2011.0378
M3 - Article
C2 - 22050430
AN - SCOPUS:84863233246
VL - 10
SP - 61
EP - 68
JO - Assay and Drug Development Technologies
JF - Assay and Drug Development Technologies
SN - 1540-658X
IS - 1
ER -