Recombinant SARS-CoV-2 Spike Protein Expression

The recombinant variant-design Spike protein of SARS-CoV-2 was expressed in the CHO-K1 cell line. The CHO-K1 cell line selected for this product came from European Collection of Authenticated Cell Cultures (ECACC) (No. 85051005). Zhongshan Kangtian Shenghe Biotechnology Co., Ltd., QuaCell, one of our partners, obtained the original adherent CHO-K1 cell from Public Health England (PHE) in England. The cells were domesticated in serum-free and suspension culture (protected by patent application) by them, and the cell lines have also been used in previous publications [18 (link), 19 (link)]. Glu-C and trypsin, both sequencing grade, were purchased from Roche and Sigma-Aldrich, respectively. Other reagents, such as peptide-N-glycosidase F (PNGase F, New England Biolabs), iodiacetamide (IAM, Sigma-Aldrich), guanidine hydrochloride (Gu.HCl, Invitrogen), and DL-dithiothreitol (DTT, VWR) were all from commercial sources.

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Huang J., Hou S., An J, & Zhou C. (2023). In-depth characterization of protein N-glycosylation for a COVID-19 variant-design vaccine spike protein. Analytical and Bioanalytical Chemistry, 415(8), 1455-1464.

Publication 2023

Glu-C trypsin peptide-N-glycosidase F (PNGase F, guanidine hydrochloride (Gu.HCl,

Cell cultures Cell lines Cells Cho cell Dithiothreitol European Guanidine hydrochloride Peptide n glycosidase f Sars cov 2 spike protein Serum Shenghe Trypsin

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Variable analysis

independent variables

Expression of the recombinant variant-design Spike protein of SARS-CoV-2 in the CHO-K1 cell line

dependent variables

Not explicitly mentioned

control variables

CHO-K1 cell line obtained from the European Collection of Authenticated Cell Cultures (ECACC) (No. 85051005)
CHO-K1 cell line domesticated in serum-free and suspension culture by Zhongshan Kangtian Shenghe Biotechnology Co., Ltd., QuaCell
Use of Glu-C and trypsin, both sequencing grade, purchased from Roche and Sigma-Aldrich, respectively
Use of other reagents, such as peptide-N-glycosidase F (PNGase F, New England Biolabs), iodiacetamide (IAM, Sigma-Aldrich), guanidine hydrochloride (Gu.HCl, Invitrogen), and DL-dithiothreitol (DTT, VWR), all from commercial sources

controls

No positive or negative controls were explicitly mentioned in the provided information.

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