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Evaluation of bioluminescence-based assays of anti-malarial drug activity

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 lu...

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Autores principales: Hasenkamp, Sandra, Sidaway, Adam, Devine, Oliver, Roye, Richard, Horrocks, Paul
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3571881/
https://www.ncbi.nlm.nih.gov/pubmed/23394077
http://dx.doi.org/10.1186/1475-2875-12-58
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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
collection PubMed
description 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.
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spelling pubmed-35718812013-02-14 Evaluation of bioluminescence-based assays of anti-malarial drug activity Hasenkamp, Sandra Sidaway, Adam Devine, Oliver Roye, Richard Horrocks, Paul Malar J Research 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. BioMed Central 2013-02-08 /pmc/articles/PMC3571881/ /pubmed/23394077 http://dx.doi.org/10.1186/1475-2875-12-58 Text en Copyright ©2013 Hasenkamp et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Hasenkamp, Sandra
Sidaway, Adam
Devine, Oliver
Roye, Richard
Horrocks, Paul
Evaluation of bioluminescence-based assays of anti-malarial drug activity
title 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_short Evaluation of bioluminescence-based assays of anti-malarial drug activity
title_sort evaluation of bioluminescence-based assays of anti-malarial drug activity
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3571881/
https://www.ncbi.nlm.nih.gov/pubmed/23394077
http://dx.doi.org/10.1186/1475-2875-12-58
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