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An inter-laboratory comparison of standard membrane-feeding assays for evaluation of malaria transmission-blocking vaccines
BACKGROUND: An effective malaria transmission-blocking vaccine may play an important role in malaria elimination efforts, and a robust biological assay is essential for its development. The standard membrane-feeding assay (SMFA) for Plasmodium falciparum infection of mosquitoes is considered a “gold...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5016893/ https://www.ncbi.nlm.nih.gov/pubmed/27612458 http://dx.doi.org/10.1186/s12936-016-1515-z |
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author | Miura, Kazutoyo Stone, Will J. R. Koolen, Karin M. Deng, Bingbing Zhou, Luwen van Gemert, Geert-Jan Locke, Emily Morin, Merribeth Bousema, Teun Sauerwein, Robert W. Long, Carole A. Dechering, Koen J. |
author_facet | Miura, Kazutoyo Stone, Will J. R. Koolen, Karin M. Deng, Bingbing Zhou, Luwen van Gemert, Geert-Jan Locke, Emily Morin, Merribeth Bousema, Teun Sauerwein, Robert W. Long, Carole A. Dechering, Koen J. |
author_sort | Miura, Kazutoyo |
collection | PubMed |
description | BACKGROUND: An effective malaria transmission-blocking vaccine may play an important role in malaria elimination efforts, and a robust biological assay is essential for its development. The standard membrane-feeding assay (SMFA) for Plasmodium falciparum infection of mosquitoes is considered a “gold standard” assay to measure transmission-blocking activity of test antibodies, and has been utilized widely in both non-clinical and clinical studies. While several studies have discussed the inherent variability of SMFA within a study group, there has been no assessment of inter-laboratory variation. Therefore, there is currently no assurance that SMFA results are comparable between different studies. METHODS: Mouse anti-Pfs25 monoclonal antibody (mAb, 4B7 mAb), rat anti-Pfs48/45 mAb (85RF45.1 mAb) and a human polyclonal antibody (pAb) collected from a malaria-exposed adult were tested at the same concentrations (6–94 μg/mL for 4B7, 1.2–31.3 μg/mL for 85RF45.1 and 23–630 μg/mL for human pAb) in two laboratories following their own standardized SMFA protocols. The mAbs and pAb, previously shown to have strong inhibition activities in the SMFA, were tested at three or four concentrations in two or three independent assays in each laboratory, and percent inhibition in mean oocyst intensity relative to a control in the same feed was determined in each feeding experiment. RESULTS: Both monoclonal and polyclonal antibodies dose-dependently reduced oocyst intensity in all experiments performed at the two test sites. In both laboratories, the inter-assay variability in percent inhibition in oocyst intensity decreased at higher levels of inhibition, regardless of which antibody was tested. At antibody concentrations that led to a >80 % reduction in oocyst numbers, the inter-laboratory variations were in the same range compared with the inter-assay variation observed within a single laboratory, and the differences in best estimates from multiple feeds between the two laboratories were <5 percentage points. CONCLUSIONS: This study confirms previous reports that the precision of the SMFA increases with increasing percent inhibition. Moreover, the variation between the two laboratories is not greater than the variation observed within a laboratory. The findings of this study provide guidance for comparison of SMFA data from different laboratories. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12936-016-1515-z) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-5016893 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-50168932016-09-10 An inter-laboratory comparison of standard membrane-feeding assays for evaluation of malaria transmission-blocking vaccines Miura, Kazutoyo Stone, Will J. R. Koolen, Karin M. Deng, Bingbing Zhou, Luwen van Gemert, Geert-Jan Locke, Emily Morin, Merribeth Bousema, Teun Sauerwein, Robert W. Long, Carole A. Dechering, Koen J. Malar J Research BACKGROUND: An effective malaria transmission-blocking vaccine may play an important role in malaria elimination efforts, and a robust biological assay is essential for its development. The standard membrane-feeding assay (SMFA) for Plasmodium falciparum infection of mosquitoes is considered a “gold standard” assay to measure transmission-blocking activity of test antibodies, and has been utilized widely in both non-clinical and clinical studies. While several studies have discussed the inherent variability of SMFA within a study group, there has been no assessment of inter-laboratory variation. Therefore, there is currently no assurance that SMFA results are comparable between different studies. METHODS: Mouse anti-Pfs25 monoclonal antibody (mAb, 4B7 mAb), rat anti-Pfs48/45 mAb (85RF45.1 mAb) and a human polyclonal antibody (pAb) collected from a malaria-exposed adult were tested at the same concentrations (6–94 μg/mL for 4B7, 1.2–31.3 μg/mL for 85RF45.1 and 23–630 μg/mL for human pAb) in two laboratories following their own standardized SMFA protocols. The mAbs and pAb, previously shown to have strong inhibition activities in the SMFA, were tested at three or four concentrations in two or three independent assays in each laboratory, and percent inhibition in mean oocyst intensity relative to a control in the same feed was determined in each feeding experiment. RESULTS: Both monoclonal and polyclonal antibodies dose-dependently reduced oocyst intensity in all experiments performed at the two test sites. In both laboratories, the inter-assay variability in percent inhibition in oocyst intensity decreased at higher levels of inhibition, regardless of which antibody was tested. At antibody concentrations that led to a >80 % reduction in oocyst numbers, the inter-laboratory variations were in the same range compared with the inter-assay variation observed within a single laboratory, and the differences in best estimates from multiple feeds between the two laboratories were <5 percentage points. CONCLUSIONS: This study confirms previous reports that the precision of the SMFA increases with increasing percent inhibition. Moreover, the variation between the two laboratories is not greater than the variation observed within a laboratory. The findings of this study provide guidance for comparison of SMFA data from different laboratories. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12936-016-1515-z) contains supplementary material, which is available to authorized users. BioMed Central 2016-09-09 /pmc/articles/PMC5016893/ /pubmed/27612458 http://dx.doi.org/10.1186/s12936-016-1515-z Text en © The Author(s) 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Miura, Kazutoyo Stone, Will J. R. Koolen, Karin M. Deng, Bingbing Zhou, Luwen van Gemert, Geert-Jan Locke, Emily Morin, Merribeth Bousema, Teun Sauerwein, Robert W. Long, Carole A. Dechering, Koen J. An inter-laboratory comparison of standard membrane-feeding assays for evaluation of malaria transmission-blocking vaccines |
title | An inter-laboratory comparison of standard membrane-feeding assays for evaluation of malaria transmission-blocking vaccines |
title_full | An inter-laboratory comparison of standard membrane-feeding assays for evaluation of malaria transmission-blocking vaccines |
title_fullStr | An inter-laboratory comparison of standard membrane-feeding assays for evaluation of malaria transmission-blocking vaccines |
title_full_unstemmed | An inter-laboratory comparison of standard membrane-feeding assays for evaluation of malaria transmission-blocking vaccines |
title_short | An inter-laboratory comparison of standard membrane-feeding assays for evaluation of malaria transmission-blocking vaccines |
title_sort | inter-laboratory comparison of standard membrane-feeding assays for evaluation of malaria transmission-blocking vaccines |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5016893/ https://www.ncbi.nlm.nih.gov/pubmed/27612458 http://dx.doi.org/10.1186/s12936-016-1515-z |
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