Evaluation of bioluminescence-based assays of anti-malarial drug activity
Abstract Background Transgenic Plasmodium falciparum expressing luciferase offers an attractive bioluminescence-based assay platform for the investigation of the pharmacological properties of anti-malarial drugs. Here a side-by-side comparison of bioluminescence and fluorescence-based assays, utiliz...
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ftdoajarticles:oai:doaj.org/article:fe5c451e829644c68de55f80e516b702 2023-05-15T15:13:31+02:00 Evaluation of bioluminescence-based assays of anti-malarial drug activity Hasenkamp Sandra Sidaway Adam Devine Oliver Roye Richard Horrocks Paul 2013-02-01T00:00:00Z https://doi.org/10.1186/1475-2875-12-58 https://doaj.org/article/fe5c451e829644c68de55f80e516b702 EN eng BMC http://www.malariajournal.com/content/12/1/58 https://doaj.org/toc/1475-2875 doi:10.1186/1475-2875-12-58 1475-2875 https://doaj.org/article/fe5c451e829644c68de55f80e516b702 Malaria Journal, Vol 12, Iss 1, p 58 (2013) Luciferase Sybr green I Antimalarial drug Relative rate of kill Malaria Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 2013 ftdoajarticles https://doi.org/10.1186/1475-2875-12-58 2022-12-31T11:52:07Z Abstract Background Transgenic Plasmodium falciparum expressing luciferase offers an attractive bioluminescence-based assay platform for the investigation of the pharmacological properties of anti-malarial drugs. Here a side-by-side comparison of bioluminescence and fluorescence-based assays, utilizing a luciferase reporter cassette that confers a strong temporal pattern of luciferase expression during the S-phase of intraerythrocytic development, is reported. Methods Key assay parameters for a range of commercially available luminogenic substrates are determined and compared to those measured using a Malaria Sybr Green I fluorescence assay. In addition, the short-term temporal effects of anti-malarial compounds are evaluated using both bioluminescent and fluorescent assay platforms. Results The Z’, % coefficient of variation and 50% inhibition concentrations are essentially the same for bioluminescent and fluorescent assays in transgenic parasites generated in both chloroquine-sensitive and -resistant genetic backgrounds. Bioluminescent assays, irrespective of the luminogenic agent employed, do, however, offer significantly enhanced signal-to-noise ratios. Moreover, the bioluminescent assay is more dynamic in terms of determining temporal effects immediately following drug perturbation. Conclusion This study suggests that opportunities for bioluminescence-based assays lie not in the measurement of 50% inhibition concentrations, where the cheaper fluorescence assay performs excellently and is not restricted by the need to genetically modify the parasite clone under investigation. Instead, assays that use the dynamic response of the luciferase reporter for semi-automated screening of additional pharmacological properties, such as relative rate-of-kill and lethal dose estimation, are a more attractive development opportunity. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Malaria Journal 12 1 58 |
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Open Polar |
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Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Luciferase Sybr green I Antimalarial drug Relative rate of kill Malaria Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
spellingShingle |
Luciferase Sybr green I Antimalarial drug Relative rate of kill Malaria Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 Hasenkamp Sandra Sidaway Adam Devine Oliver Roye Richard Horrocks Paul Evaluation of bioluminescence-based assays of anti-malarial drug activity |
topic_facet |
Luciferase Sybr green I Antimalarial drug Relative rate of kill Malaria Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
Abstract Background Transgenic Plasmodium falciparum expressing luciferase offers an attractive bioluminescence-based assay platform for the investigation of the pharmacological properties of anti-malarial drugs. Here a side-by-side comparison of bioluminescence and fluorescence-based assays, utilizing a luciferase reporter cassette that confers a strong temporal pattern of luciferase expression during the S-phase of intraerythrocytic development, is reported. Methods Key assay parameters for a range of commercially available luminogenic substrates are determined and compared to those measured using a Malaria Sybr Green I fluorescence assay. In addition, the short-term temporal effects of anti-malarial compounds are evaluated using both bioluminescent and fluorescent assay platforms. Results The Z’, % coefficient of variation and 50% inhibition concentrations are essentially the same for bioluminescent and fluorescent assays in transgenic parasites generated in both chloroquine-sensitive and -resistant genetic backgrounds. Bioluminescent assays, irrespective of the luminogenic agent employed, do, however, offer significantly enhanced signal-to-noise ratios. Moreover, the bioluminescent assay is more dynamic in terms of determining temporal effects immediately following drug perturbation. Conclusion This study suggests that opportunities for bioluminescence-based assays lie not in the measurement of 50% inhibition concentrations, where the cheaper fluorescence assay performs excellently and is not restricted by the need to genetically modify the parasite clone under investigation. Instead, assays that use the dynamic response of the luciferase reporter for semi-automated screening of additional pharmacological properties, such as relative rate-of-kill and lethal dose estimation, are a more attractive development opportunity. |
format |
Article in Journal/Newspaper |
author |
Hasenkamp Sandra Sidaway Adam Devine Oliver Roye Richard Horrocks Paul |
author_facet |
Hasenkamp Sandra Sidaway Adam Devine Oliver Roye Richard Horrocks Paul |
author_sort |
Hasenkamp Sandra |
title |
Evaluation of bioluminescence-based assays of anti-malarial drug activity |
title_short |
Evaluation of bioluminescence-based assays of anti-malarial drug activity |
title_full |
Evaluation of bioluminescence-based assays of anti-malarial drug activity |
title_fullStr |
Evaluation of bioluminescence-based assays of anti-malarial drug activity |
title_full_unstemmed |
Evaluation of bioluminescence-based assays of anti-malarial drug activity |
title_sort |
evaluation of bioluminescence-based assays of anti-malarial drug activity |
publisher |
BMC |
publishDate |
2013 |
url |
https://doi.org/10.1186/1475-2875-12-58 https://doaj.org/article/fe5c451e829644c68de55f80e516b702 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Malaria Journal, Vol 12, Iss 1, p 58 (2013) |
op_relation |
http://www.malariajournal.com/content/12/1/58 https://doaj.org/toc/1475-2875 doi:10.1186/1475-2875-12-58 1475-2875 https://doaj.org/article/fe5c451e829644c68de55f80e516b702 |
op_doi |
https://doi.org/10.1186/1475-2875-12-58 |
container_title |
Malaria Journal |
container_volume |
12 |
container_issue |
1 |
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58 |
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1766344063803457536 |