Recombinant production of a functional SARS-CoV-2 spike receptor binding domain in the green algae Chlamydomonas reinhardtii.

Recombinant production of viral proteins can be used to produce vaccine antigens or reagents to identify antibodies in patient serum. Minimally, these proteins must be correctly folded and have appropriate post-translation modifications. Here we report the production of the SARS-CoV-2 spike protein...

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Autores principales:	Anthony J Berndt, Tressa N Smalley, Bijie Ren, Ryan Simkovsky, Amr Badary, Ashley E Sproles, Francis J Fields, Yasin Torres-Tiji, Vanessa Heredia, Stephen P Mayfield
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e852bfaff8a342708b82c8e319589b06
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spelling	oai:doaj.org-article:e852bfaff8a342708b82c8e319589b062021-12-02T20:19:10ZRecombinant production of a functional SARS-CoV-2 spike receptor binding domain in the green algae Chlamydomonas reinhardtii.1932-620310.1371/journal.pone.0257089https://doaj.org/article/e852bfaff8a342708b82c8e319589b062021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0257089https://doaj.org/toc/1932-6203Recombinant production of viral proteins can be used to produce vaccine antigens or reagents to identify antibodies in patient serum. Minimally, these proteins must be correctly folded and have appropriate post-translation modifications. Here we report the production of the SARS-CoV-2 spike protein Receptor Binding Domain (RBD) in the green algae Chlamydomonas. RBD fused to a fluorescent reporter protein accumulates as an intact protein when targeted for ER-Golgi retention or secreted from the cell, while a chloroplast localized version is truncated. The ER-retained RBD fusion protein was able to bind the human ACE2 receptor, the host target of SARS-CoV-2, and was specifically out-competed by mammalian cell-produced recombinant RBD, suggesting that the algae produced proteins are sufficiently post-translationally modified to act as authentic SARS-CoV-2 antigens. Because algae can be grown at large scale very inexpensively, this recombinant protein may be a low cost alternative to other expression platforms.Anthony J BerndtTressa N SmalleyBijie RenRyan SimkovskyAmr BadaryAshley E SprolesFrancis J FieldsYasin Torres-TijiVanessa HerediaStephen P MayfieldPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 11, p e0257089 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Anthony J Berndt Tressa N Smalley Bijie Ren Ryan Simkovsky Amr Badary Ashley E Sproles Francis J Fields Yasin Torres-Tiji Vanessa Heredia Stephen P Mayfield Recombinant production of a functional SARS-CoV-2 spike receptor binding domain in the green algae Chlamydomonas reinhardtii.
description	Recombinant production of viral proteins can be used to produce vaccine antigens or reagents to identify antibodies in patient serum. Minimally, these proteins must be correctly folded and have appropriate post-translation modifications. Here we report the production of the SARS-CoV-2 spike protein Receptor Binding Domain (RBD) in the green algae Chlamydomonas. RBD fused to a fluorescent reporter protein accumulates as an intact protein when targeted for ER-Golgi retention or secreted from the cell, while a chloroplast localized version is truncated. The ER-retained RBD fusion protein was able to bind the human ACE2 receptor, the host target of SARS-CoV-2, and was specifically out-competed by mammalian cell-produced recombinant RBD, suggesting that the algae produced proteins are sufficiently post-translationally modified to act as authentic SARS-CoV-2 antigens. Because algae can be grown at large scale very inexpensively, this recombinant protein may be a low cost alternative to other expression platforms.
format	article
author	Anthony J Berndt Tressa N Smalley Bijie Ren Ryan Simkovsky Amr Badary Ashley E Sproles Francis J Fields Yasin Torres-Tiji Vanessa Heredia Stephen P Mayfield
author_facet	Anthony J Berndt Tressa N Smalley Bijie Ren Ryan Simkovsky Amr Badary Ashley E Sproles Francis J Fields Yasin Torres-Tiji Vanessa Heredia Stephen P Mayfield
author_sort	Anthony J Berndt
title	Recombinant production of a functional SARS-CoV-2 spike receptor binding domain in the green algae Chlamydomonas reinhardtii.
title_short	Recombinant production of a functional SARS-CoV-2 spike receptor binding domain in the green algae Chlamydomonas reinhardtii.
title_full	Recombinant production of a functional SARS-CoV-2 spike receptor binding domain in the green algae Chlamydomonas reinhardtii.
title_fullStr	Recombinant production of a functional SARS-CoV-2 spike receptor binding domain in the green algae Chlamydomonas reinhardtii.
title_full_unstemmed	Recombinant production of a functional SARS-CoV-2 spike receptor binding domain in the green algae Chlamydomonas reinhardtii.
title_sort	recombinant production of a functional sars-cov-2 spike receptor binding domain in the green algae chlamydomonas reinhardtii.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/e852bfaff8a342708b82c8e319589b06
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Recombinant production of a functional SARS-CoV-2 spike receptor binding domain in the green algae Chlamydomonas reinhardtii.

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