Production and secretion of functional SARS-CoV-2 spike protein in Chlamydomonas reinhardtii
The spike protein is the major protein on the surface of coronaviruses. It is therefore the prominent target of neutralizing antibodies and consequently the antigen of all currently admitted vaccines against SARS-CoV-2. Since it is a 1,273-amino acids glycoprotein with 22 N-linked glycans, the produ...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-09-01
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| Series: | Frontiers in Plant Science |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fpls.2022.988870/full |
| _version_ | 1811266479751954432 |
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| author | Anna Maria Kiefer Justus Niemeyer Anna Probst Gerhard Erkel Michael Schroda |
| author_facet | Anna Maria Kiefer Justus Niemeyer Anna Probst Gerhard Erkel Michael Schroda |
| author_sort | Anna Maria Kiefer |
| collection | DOAJ |
| description | The spike protein is the major protein on the surface of coronaviruses. It is therefore the prominent target of neutralizing antibodies and consequently the antigen of all currently admitted vaccines against SARS-CoV-2. Since it is a 1,273-amino acids glycoprotein with 22 N-linked glycans, the production of functional, full-length spike protein was limited to higher eukaryotes. Here we report the production of full-length SARS-CoV-2 spike protein – lacking the C-terminal membrane anchor – as a secreted protein in the prefusion-stabilized conformation in the unicellular green alga Chlamydomonas reinhardtii. We show that the spike protein is efficiently cleaved at the furin cleavage site during synthesis in the alga and that cleavage is abolished upon mutation of the multi-basic cleavage site. We could enrich the spike protein from culture medium by ammonium sulfate precipitation and demonstrate its functionality based on its interaction with recombinant ACE2 and ACE2 expressed on human 293T cells. Chlamydomonas reinhardtii is a GRAS organism that can be cultivated at low cost in simple media at a large scale, making it an attractive production platform for recombinant spike protein and other biopharmaceuticals in low-income countries. |
| first_indexed | 2024-04-12T20:42:51Z |
| format | Article |
| id | doaj.art-b0b070d789be42049a64d8a582ff3e1d |
| institution | Directory Open Access Journal |
| issn | 1664-462X |
| language | English |
| last_indexed | 2024-04-12T20:42:51Z |
| publishDate | 2022-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Plant Science |
| spelling | doaj.art-b0b070d789be42049a64d8a582ff3e1d2022-12-22T03:17:21ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2022-09-011310.3389/fpls.2022.988870988870Production and secretion of functional SARS-CoV-2 spike protein in Chlamydomonas reinhardtiiAnna Maria KieferJustus NiemeyerAnna ProbstGerhard ErkelMichael SchrodaThe spike protein is the major protein on the surface of coronaviruses. It is therefore the prominent target of neutralizing antibodies and consequently the antigen of all currently admitted vaccines against SARS-CoV-2. Since it is a 1,273-amino acids glycoprotein with 22 N-linked glycans, the production of functional, full-length spike protein was limited to higher eukaryotes. Here we report the production of full-length SARS-CoV-2 spike protein – lacking the C-terminal membrane anchor – as a secreted protein in the prefusion-stabilized conformation in the unicellular green alga Chlamydomonas reinhardtii. We show that the spike protein is efficiently cleaved at the furin cleavage site during synthesis in the alga and that cleavage is abolished upon mutation of the multi-basic cleavage site. We could enrich the spike protein from culture medium by ammonium sulfate precipitation and demonstrate its functionality based on its interaction with recombinant ACE2 and ACE2 expressed on human 293T cells. Chlamydomonas reinhardtii is a GRAS organism that can be cultivated at low cost in simple media at a large scale, making it an attractive production platform for recombinant spike protein and other biopharmaceuticals in low-income countries.https://www.frontiersin.org/articles/10.3389/fpls.2022.988870/fullsynthetic biologyglycoproteinsbiopharmaceuticalsmicroalgal production platformfurin-like protease |
| spellingShingle | Anna Maria Kiefer Justus Niemeyer Anna Probst Gerhard Erkel Michael Schroda Production and secretion of functional SARS-CoV-2 spike protein in Chlamydomonas reinhardtii Frontiers in Plant Science synthetic biology glycoproteins biopharmaceuticals microalgal production platform furin-like protease |
| title | Production and secretion of functional SARS-CoV-2 spike protein in Chlamydomonas reinhardtii |
| title_full | Production and secretion of functional SARS-CoV-2 spike protein in Chlamydomonas reinhardtii |
| title_fullStr | Production and secretion of functional SARS-CoV-2 spike protein in Chlamydomonas reinhardtii |
| title_full_unstemmed | Production and secretion of functional SARS-CoV-2 spike protein in Chlamydomonas reinhardtii |
| title_short | Production and secretion of functional SARS-CoV-2 spike protein in Chlamydomonas reinhardtii |
| title_sort | production and secretion of functional sars cov 2 spike protein in chlamydomonas reinhardtii |
| topic | synthetic biology glycoproteins biopharmaceuticals microalgal production platform furin-like protease |
| url | https://www.frontiersin.org/articles/10.3389/fpls.2022.988870/full |
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