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Production of full-length soluble Plasmodium falciparum RH5 protein vaccine using a Drosophila melanogaster Schneider 2 stable cell line system
The Plasmodium falciparum reticulocyte-binding protein homolog 5 (PfRH5) has recently emerged as a leading candidate antigen against the blood-stage human malaria parasite. However it has proved challenging to identify a heterologous expression platform that can produce a soluble protein-based vacci...
Autores principales: | , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4960544/ https://www.ncbi.nlm.nih.gov/pubmed/27457156 http://dx.doi.org/10.1038/srep30357 |
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author | Hjerrild, Kathryn A. Jin, Jing Wright, Katherine E. Brown, Rebecca E. Marshall, Jennifer M. Labbé, Geneviève M. Silk, Sarah E. Cherry, Catherine J. Clemmensen, Stine B. Jørgensen, Thomas Illingworth, Joseph J. Alanine, Daniel G. W. Milne, Kathryn H. Ashfield, Rebecca de Jongh, Willem A. Douglas, Alexander D. Higgins, Matthew K. Draper, Simon J. |
author_facet | Hjerrild, Kathryn A. Jin, Jing Wright, Katherine E. Brown, Rebecca E. Marshall, Jennifer M. Labbé, Geneviève M. Silk, Sarah E. Cherry, Catherine J. Clemmensen, Stine B. Jørgensen, Thomas Illingworth, Joseph J. Alanine, Daniel G. W. Milne, Kathryn H. Ashfield, Rebecca de Jongh, Willem A. Douglas, Alexander D. Higgins, Matthew K. Draper, Simon J. |
author_sort | Hjerrild, Kathryn A. |
collection | PubMed |
description | The Plasmodium falciparum reticulocyte-binding protein homolog 5 (PfRH5) has recently emerged as a leading candidate antigen against the blood-stage human malaria parasite. However it has proved challenging to identify a heterologous expression platform that can produce a soluble protein-based vaccine in a manner compliant with current Good Manufacturing Practice (cGMP). Here we report the production of full-length PfRH5 protein using a cGMP-compliant platform called ExpreS(2), based on a Drosophila melanogaster Schneider 2 (S2) stable cell line system. Five sequence variants of PfRH5 were expressed that differed in terms of mutagenesis strategies to remove potential N-linked glycans. All variants bound the PfRH5 receptor basigin and were recognized by a panel of monoclonal antibodies. Analysis following immunization of rabbits identified quantitative and qualitative differences in terms of the functional IgG antibody response against the P. falciparum parasite. The antibodies induced by one protein variant were shown to be qualitatively similar to responses induced by other vaccine platforms. This work identifies Drosophila S2 cells as a clinically-relevant platform suited for the production of ‘difficult-to-make’ proteins from Plasmodium parasites, and identifies a PfRH5 sequence variant that can be used for clinical production of a non-glycosylated, soluble full-length protein vaccine immunogen. |
format | Online Article Text |
id | pubmed-4960544 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-49605442016-08-05 Production of full-length soluble Plasmodium falciparum RH5 protein vaccine using a Drosophila melanogaster Schneider 2 stable cell line system Hjerrild, Kathryn A. Jin, Jing Wright, Katherine E. Brown, Rebecca E. Marshall, Jennifer M. Labbé, Geneviève M. Silk, Sarah E. Cherry, Catherine J. Clemmensen, Stine B. Jørgensen, Thomas Illingworth, Joseph J. Alanine, Daniel G. W. Milne, Kathryn H. Ashfield, Rebecca de Jongh, Willem A. Douglas, Alexander D. Higgins, Matthew K. Draper, Simon J. Sci Rep Article The Plasmodium falciparum reticulocyte-binding protein homolog 5 (PfRH5) has recently emerged as a leading candidate antigen against the blood-stage human malaria parasite. However it has proved challenging to identify a heterologous expression platform that can produce a soluble protein-based vaccine in a manner compliant with current Good Manufacturing Practice (cGMP). Here we report the production of full-length PfRH5 protein using a cGMP-compliant platform called ExpreS(2), based on a Drosophila melanogaster Schneider 2 (S2) stable cell line system. Five sequence variants of PfRH5 were expressed that differed in terms of mutagenesis strategies to remove potential N-linked glycans. All variants bound the PfRH5 receptor basigin and were recognized by a panel of monoclonal antibodies. Analysis following immunization of rabbits identified quantitative and qualitative differences in terms of the functional IgG antibody response against the P. falciparum parasite. The antibodies induced by one protein variant were shown to be qualitatively similar to responses induced by other vaccine platforms. This work identifies Drosophila S2 cells as a clinically-relevant platform suited for the production of ‘difficult-to-make’ proteins from Plasmodium parasites, and identifies a PfRH5 sequence variant that can be used for clinical production of a non-glycosylated, soluble full-length protein vaccine immunogen. Nature Publishing Group 2016-07-26 /pmc/articles/PMC4960544/ /pubmed/27457156 http://dx.doi.org/10.1038/srep30357 Text en Copyright © 2016, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Hjerrild, Kathryn A. Jin, Jing Wright, Katherine E. Brown, Rebecca E. Marshall, Jennifer M. Labbé, Geneviève M. Silk, Sarah E. Cherry, Catherine J. Clemmensen, Stine B. Jørgensen, Thomas Illingworth, Joseph J. Alanine, Daniel G. W. Milne, Kathryn H. Ashfield, Rebecca de Jongh, Willem A. Douglas, Alexander D. Higgins, Matthew K. Draper, Simon J. Production of full-length soluble Plasmodium falciparum RH5 protein vaccine using a Drosophila melanogaster Schneider 2 stable cell line system |
title | Production of full-length soluble Plasmodium falciparum RH5 protein vaccine using a Drosophila melanogaster Schneider 2 stable cell line system |
title_full | Production of full-length soluble Plasmodium falciparum RH5 protein vaccine using a Drosophila melanogaster Schneider 2 stable cell line system |
title_fullStr | Production of full-length soluble Plasmodium falciparum RH5 protein vaccine using a Drosophila melanogaster Schneider 2 stable cell line system |
title_full_unstemmed | Production of full-length soluble Plasmodium falciparum RH5 protein vaccine using a Drosophila melanogaster Schneider 2 stable cell line system |
title_short | Production of full-length soluble Plasmodium falciparum RH5 protein vaccine using a Drosophila melanogaster Schneider 2 stable cell line system |
title_sort | production of full-length soluble plasmodium falciparum rh5 protein vaccine using a drosophila melanogaster schneider 2 stable cell line system |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4960544/ https://www.ncbi.nlm.nih.gov/pubmed/27457156 http://dx.doi.org/10.1038/srep30357 |
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