Recombinant SARS-CoV-2, MERS-CoV, and Endemic Coronavirus Proteins

Recombinant SARS-CoV-2 S1 subunit of the S protein (amino acids 1–685, expressed in mammalian HEK293 cells), MERS-CoV S1 subunit (amino acids 1–725, expressed in mammalian HEK293 cells), and full-length S proteins (expressed in baculovirus-insect cells) from hCoV-OC43, hCoV-NL63, hCoV-229E, and hCoV-HKU1 viruses tagged with histidine tag (His-tag) were purchased commercially (Sino Biological, China). Recombinant SARS-CoV-2 and MERS-CoV N proteins were expressed and purified from Escherichia coli BL21 (DE3) cells using a nickel-nitrilotriacetic acid (Ni-NTA) column according to the manufacturer's protocol and as previously described^{7 (link)}. Positive fractions of N proteins were pooled, aliquoted, and stored at − 80 °C until used. SARS-CoV-2 proteins were confirmed by Western blot using anti-His tag antibodies as well as SARS-CoV-2 seropositive and seronegative human serum samples as previously described^{7 (link)}.

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Algaissi A., Alfaleh M.A., Hala S., Abujamel T.S., Alamri S.S., Almahboub S.A., Alluhaybi K.A., Hobani H.I., Alsulaiman R.M., AlHarbi R.H., ElAssouli M.Z., Alhabbab R.Y., AlSaieedi A.A., Abdulaal W.H., Al-Somali A.A., Alofi F.S., Khogeer A.A., Alkayyal A.A., Mahmoud A.B., Almontashiri N.A., Pain A, & Hashem A.M. (2020). SARS-CoV-2 S1 and N-based serological assays reveal rapid seroconversion and induction of specific antibody response in COVID-19 patients. Scientific Reports, 10, 16561.

Publication 2020

hCoV-OC43 hCoV-NL63 hCoV-229E hCoV-HKU1

Amino acids Antibodies Baculovirus Biological Cells Escherichia coli Hcov 229e Hcov hku1 Hcov nl63 Hcov oc43 Hek293 cells Histidine Human Insect Mammalian Mers cov Nickel nitrilotriacetic acid Proteins Proteins n Sars cov 2 Sars cov 2 s protein Serum Sino Subunit Viruses Western blot

Corresponding Organization : King Abdulaziz University

Other organizations : King Abdullah University of Science and Technology, King Fahad Hospital Jeddah, Hera General Hospital, University of Tabuk, Taibah University

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Variable analysis

independent variables

Recombinant SARS-CoV-2 S1 subunit of the S protein (amino acids 1–685, expressed in mammalian HEK293 cells)
MERS-CoV S1 subunit (amino acids 1–725, expressed in mammalian HEK293 cells)
Full-length S proteins (expressed in baculovirus-insect cells) from hCoV-OC43, hCoV-NL63, hCoV-229E, and hCoV-HKU1 viruses
Recombinant SARS-CoV-2 and MERS-CoV N proteins expressed and purified from Escherichia coli BL21 (DE3) cells

dependent variables

Not explicitly mentioned

control variables

His-tag used to tag the viral proteins
Positive and negative human serum samples used to confirm SARS-CoV-2 proteins

positive controls

SARS-CoV-2 seropositive human serum samples

negative controls

SARS-CoV-2 seronegative human serum samples

Annotations

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