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Plant production of recombinant antigens containing the receptor binding domain (RBD) of two SARS-CoV-2 variants.
Plant production of recombinant antigens containing the receptor binding domain (RBD) of two SARS-CoV-2 variants.
- Source :
-
Biotechnology letters [Biotechnol Lett] 2024 Dec; Vol. 46 (6), pp. 1303-1318. Date of Electronic Publication: 2024 Jul 27. - Publication Year :
- 2024
-
Abstract
- Objectives: The aim of this work was to rapidly produce in plats two recombinant antigens (RBDw-Fc and RBDo-Fc) containing the receptor binding domain (RBD) of the spike (S) protein from SARS-CoV-2 variants Wuhan and Omicron as fusion proteins to the Fc portion of a murine IgG2a antibody constant region (Fc).<br />Results: The two recombinant antigens were expressed in Nicotiana benthamiana plants, engineered to avoid the addition of N-linked plant-typical sugars, through vacuum agroinfiltration and showed comparable purification yields (about 35 mg/kg leaf fresh weight).<br />Conclusions: Their Western blotting and Coomassie staining evidenced the occurrence of major in planta proteolysis in the region between the RBD and Fc, which was particularly evident in RBDw-Fc, the only antigen bearing the HRV 3C cysteine protease recognition site. The two RBD N-linked glycosylation sites showed very homogeneous profiles free from plant-typical sugars, with the most abundant glycoform represented by the complex sugar GlcNAc <subscript>4</subscript> Man <subscript>3</subscript> . Both antigens were specifically recognised in Western Blot analysis by the anti-SARS-CoV-2 human neutralizing monoclonal antibody J08-MUT and RBDw-Fc was successfully used in competitive ELISA experiments for binding to the angiotensin-converting enzyme 2 receptor to verify the neutralizing capacity of the serum from vaccinated patients. Both SARS-Cov-2 antigens fused to a murine Fc region were rapidly and functionally produced in plants with potential applications in diagnostics.<br /> (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)
- Subjects :
- Humans
Recombinant Fusion Proteins genetics
Recombinant Fusion Proteins metabolism
Recombinant Fusion Proteins chemistry
Recombinant Fusion Proteins immunology
Animals
Antigens, Viral genetics
Antigens, Viral immunology
COVID-19
Mice
Antibodies, Viral immunology
Protein Domains
Antibodies, Neutralizing immunology
Plants, Genetically Modified genetics
Plants, Genetically Modified metabolism
Recombinant Proteins genetics
Recombinant Proteins metabolism
Recombinant Proteins chemistry
Recombinant Proteins immunology
Nicotiana genetics
Nicotiana metabolism
Spike Glycoprotein, Coronavirus genetics
Spike Glycoprotein, Coronavirus immunology
Spike Glycoprotein, Coronavirus metabolism
Spike Glycoprotein, Coronavirus chemistry
SARS-CoV-2 genetics
SARS-CoV-2 immunology
SARS-CoV-2 metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1573-6776
- Volume :
- 46
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Biotechnology letters
- Publication Type :
- Academic Journal
- Accession number :
- 39066957
- Full Text :
- https://doi.org/10.1007/s10529-024-03517-9