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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.

Authors :
Fagiani F
Frigerio R
Salzano AM
Scaloni A
Marusic C
Donini M
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.)

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