Optimization of incubation conditions of Plasmodium falciparum antibody multiplex assays to measure IgG, IgG1–4, IgM and IgE using standard and customized reference pools for sero-epidemiological and vaccine studies
Abstract Background The quantitative suspension array technology (qSAT) is a useful platform for malaria immune marker discovery. However, a major challenge for large sero-epidemiological and malaria vaccine studies is the comparability across laboratories, which requires the access to standardized...
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| Format: | Article |
| Language: | English |
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BMC
2018-06-01
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| Series: | Malaria Journal |
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| Online Access: | http://link.springer.com/article/10.1186/s12936-018-2369-3 |
| _version_ | 1819261317077270528 |
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| author | Itziar Ubillos Alfons Jiménez Marta Vidal Paul W. Bowyer Deepak Gaur Sheetij Dutta Benoit Gamain Ross Coppel Virander Chauhan David Lanar Chetan Chitnis Evelina Angov James Beeson David Cavanagh Joseph J. Campo Ruth Aguilar Carlota Dobaño |
| author_facet | Itziar Ubillos Alfons Jiménez Marta Vidal Paul W. Bowyer Deepak Gaur Sheetij Dutta Benoit Gamain Ross Coppel Virander Chauhan David Lanar Chetan Chitnis Evelina Angov James Beeson David Cavanagh Joseph J. Campo Ruth Aguilar Carlota Dobaño |
| author_sort | Itziar Ubillos |
| collection | DOAJ |
| description | Abstract Background The quantitative suspension array technology (qSAT) is a useful platform for malaria immune marker discovery. However, a major challenge for large sero-epidemiological and malaria vaccine studies is the comparability across laboratories, which requires the access to standardized control reagents for assay optimization, to monitor performance and improve reproducibility. Here, the Plasmodium falciparum antibody reactivities of the newly available WHO reference reagent for anti-malaria human plasma (10/198) and of additional customized positive controls were examined with seven in-house qSAT multiplex assays measuring IgG, IgG1–4 subclasses, IgM and IgE against a panel of 40 antigens. The different positive controls were tested at different incubation times and temperatures (4 °C overnight, 37 °C 2 h, room temperature 1 h) to select the optimal conditions. Results Overall, the WHO reference reagent had low IgG2, IgG4, IgM and IgE, and also low anti-CSP antibody levels, thus this reagent was enriched with plasmas from RTS,S-vaccinated volunteers to be used as standard for CSP-based vaccine studies. For the IgM assay, another customized plasma pool prepared with samples from malaria primo-infected adults with adequate IgM levels proved to be more adequate as a positive control. The range and magnitude of IgG and IgG1–4 responses were highest when the WHO reference reagent was incubated with antigen-coupled beads at 4 °C overnight. IgG levels measured in the negative control did not vary between incubations at 37 °C 2 h and 4 °C overnight, indicating no difference in unspecific binding. Conclusions With this study, the immunogenicity profile of the WHO reference reagent, including seven immunoglobulin isotypes and subclasses, and more P. falciparum antigens, also those included in the leading RTS,S malaria vaccine, was better characterized. Overall, incubation of samples at 4 °C overnight rendered the best performance for antibody measurements against the antigens tested. Although the WHO reference reagent performed well to measure IgG to the majority of the common P. falciparum blood stage antigens tested, customized pools may need to be used as positive controls depending on the antigens (e.g. pre-erythrocytic proteins of low natural immunogenicity) and isotypes/subclasses (e.g. IgM) under study. |
| first_indexed | 2024-12-23T19:39:52Z |
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| institution | Directory Open Access Journal |
| issn | 1475-2875 |
| language | English |
| last_indexed | 2024-12-23T19:39:52Z |
| publishDate | 2018-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Malaria Journal |
| spelling | doaj.art-5ba8377aef2545d38add042e560f87382022-12-21T17:33:42ZengBMCMalaria Journal1475-28752018-06-0117111510.1186/s12936-018-2369-3Optimization of incubation conditions of Plasmodium falciparum antibody multiplex assays to measure IgG, IgG1–4, IgM and IgE using standard and customized reference pools for sero-epidemiological and vaccine studiesItziar Ubillos0Alfons Jiménez1Marta Vidal2Paul W. Bowyer3Deepak Gaur4Sheetij Dutta5Benoit Gamain6Ross Coppel7Virander Chauhan8David Lanar9Chetan Chitnis10Evelina Angov11James Beeson12David Cavanagh13Joseph J. Campo14Ruth Aguilar15Carlota Dobaño16ISGlobal, Hospital Clínic-Universitat de BarcelonaISGlobal, Hospital Clínic-Universitat de BarcelonaISGlobal, Hospital Clínic-Universitat de BarcelonaBacteriology Division, MHRA-NIBSCLaboratory of Malaria and Vaccine Research, School of Biotechnology, Jawaharlal Nehru UniversityU.S. Military Malaria Vaccine Program, Walter Reed Army Institute of ResearchUniversité Sorbonne Paris Cité, Université Paris Diderot, Inserm, INTS, Unité Biologie Intégrée du Globule Rouge UMR_S1134, Laboratoire d’Excellence GR-ExInfection and Immunity Program, Monash Biomedicine Discovery Institute and Department of Microbiology, Monash UniversityMalaria Group, International Centre for Genetic Engineering and Biotechnology (ICGEB)U.S. Military Malaria Vaccine Program, Walter Reed Army Institute of ResearchMalaria Group, International Centre for Genetic Engineering and Biotechnology (ICGEB)U.S. Military Malaria Vaccine Program, Walter Reed Army Institute of ResearchMacfarlane Burnet Institute for Medical Research and Public HealthInstitute of Immunology & Infection Research and Centre for Immunity, Infection & Evolution, Ashworth Laboratories, School of Biological Sciences, University of EdinburghISGlobal, Hospital Clínic-Universitat de BarcelonaISGlobal, Hospital Clínic-Universitat de BarcelonaISGlobal, Hospital Clínic-Universitat de BarcelonaAbstract Background The quantitative suspension array technology (qSAT) is a useful platform for malaria immune marker discovery. However, a major challenge for large sero-epidemiological and malaria vaccine studies is the comparability across laboratories, which requires the access to standardized control reagents for assay optimization, to monitor performance and improve reproducibility. Here, the Plasmodium falciparum antibody reactivities of the newly available WHO reference reagent for anti-malaria human plasma (10/198) and of additional customized positive controls were examined with seven in-house qSAT multiplex assays measuring IgG, IgG1–4 subclasses, IgM and IgE against a panel of 40 antigens. The different positive controls were tested at different incubation times and temperatures (4 °C overnight, 37 °C 2 h, room temperature 1 h) to select the optimal conditions. Results Overall, the WHO reference reagent had low IgG2, IgG4, IgM and IgE, and also low anti-CSP antibody levels, thus this reagent was enriched with plasmas from RTS,S-vaccinated volunteers to be used as standard for CSP-based vaccine studies. For the IgM assay, another customized plasma pool prepared with samples from malaria primo-infected adults with adequate IgM levels proved to be more adequate as a positive control. The range and magnitude of IgG and IgG1–4 responses were highest when the WHO reference reagent was incubated with antigen-coupled beads at 4 °C overnight. IgG levels measured in the negative control did not vary between incubations at 37 °C 2 h and 4 °C overnight, indicating no difference in unspecific binding. Conclusions With this study, the immunogenicity profile of the WHO reference reagent, including seven immunoglobulin isotypes and subclasses, and more P. falciparum antigens, also those included in the leading RTS,S malaria vaccine, was better characterized. Overall, incubation of samples at 4 °C overnight rendered the best performance for antibody measurements against the antigens tested. Although the WHO reference reagent performed well to measure IgG to the majority of the common P. falciparum blood stage antigens tested, customized pools may need to be used as positive controls depending on the antigens (e.g. pre-erythrocytic proteins of low natural immunogenicity) and isotypes/subclasses (e.g. IgM) under study.http://link.springer.com/article/10.1186/s12936-018-2369-3Plasmodium falciparumQuantitative suspension array technologyMultiplexIgG, IgG1, IgG2, IgG3, IgG4 subclassesIgMIgE |
| spellingShingle | Itziar Ubillos Alfons Jiménez Marta Vidal Paul W. Bowyer Deepak Gaur Sheetij Dutta Benoit Gamain Ross Coppel Virander Chauhan David Lanar Chetan Chitnis Evelina Angov James Beeson David Cavanagh Joseph J. Campo Ruth Aguilar Carlota Dobaño Optimization of incubation conditions of Plasmodium falciparum antibody multiplex assays to measure IgG, IgG1–4, IgM and IgE using standard and customized reference pools for sero-epidemiological and vaccine studies Malaria Journal Plasmodium falciparum Quantitative suspension array technology Multiplex IgG, IgG1, IgG2, IgG3, IgG4 subclasses IgM IgE |
| title | Optimization of incubation conditions of Plasmodium falciparum antibody multiplex assays to measure IgG, IgG1–4, IgM and IgE using standard and customized reference pools for sero-epidemiological and vaccine studies |
| title_full | Optimization of incubation conditions of Plasmodium falciparum antibody multiplex assays to measure IgG, IgG1–4, IgM and IgE using standard and customized reference pools for sero-epidemiological and vaccine studies |
| title_fullStr | Optimization of incubation conditions of Plasmodium falciparum antibody multiplex assays to measure IgG, IgG1–4, IgM and IgE using standard and customized reference pools for sero-epidemiological and vaccine studies |
| title_full_unstemmed | Optimization of incubation conditions of Plasmodium falciparum antibody multiplex assays to measure IgG, IgG1–4, IgM and IgE using standard and customized reference pools for sero-epidemiological and vaccine studies |
| title_short | Optimization of incubation conditions of Plasmodium falciparum antibody multiplex assays to measure IgG, IgG1–4, IgM and IgE using standard and customized reference pools for sero-epidemiological and vaccine studies |
| title_sort | optimization of incubation conditions of plasmodium falciparum antibody multiplex assays to measure igg igg1 4 igm and ige using standard and customized reference pools for sero epidemiological and vaccine studies |
| topic | Plasmodium falciparum Quantitative suspension array technology Multiplex IgG, IgG1, IgG2, IgG3, IgG4 subclasses IgM IgE |
| url | http://link.springer.com/article/10.1186/s12936-018-2369-3 |
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