About: Production of Trimeric SARS-CoV-2 Spike Protein by CHO Cells for Serological COVID-19 Testing

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About: Production of Trimeric SARS-CoV-2 Spike Protein by CHO Cells for Serological COVID-19 Testing Goto Sponge NotDistinct Permalink

An Entity of Type : schema:ScholarlyArticle, within Data Space : covidontheweb.inria.fr associated with source document(s)

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	Production of Trimeric SARS-CoV-2 Spike Protein by CHO Cells for Serological COVID-19 Testing
Creator	Tee, James, David Bhayat-Cammack, Amina Brown, Adam Dickman, Mark Jackson, Philip Jaffé, Stephen Johari, Yusuf Johnson, Abayomi Kang, Lan Maisuria, Sheetal Mozzanino, Théo Pohle, Thilo Sargur, Ravishankar Scarrott, Joseph Wong, Tuck
Source	MedRxiv
abstract	We describe scalable and cost-efficient production of full length, His-tagged SARS-CoV-2 spike glycoprotein trimer by CHO cells that can be used to detect SARS-CoV-2 antibodies in patient sera at high specificity and sensitivity. Transient production of spike in both HEK and CHO cells mediated by PEI was increased significantly (up to 10.9-fold) by a reduction in culture temperature to 32C to permit extended duration cultures. Based on these data GS-CHO pools stably producing spike trimer under the control of a strong synthetic promoter were cultured in hypothermic conditions with combinations of bioactive small molecules to increase yield of purified spike product 4.9-fold to 53 mg/L. Purification of recombinant spike by Ni-chelate affinity chromatography initially yielded a variety of co-eluting protein impurities identified as host cell-derived by mass spectrometry, which were separated from spike trimer using a modified imidazole gradient elution. Purified CHO spike trimer antigen was used in ELISA format to detect IgG antibodies against SARS-CoV-2 in sera from patient cohorts previously tested for viral infection by PCR, including those who had displayed COVID-19 symptoms. The antibody assay, validated to ISO 15189 Medical Laboratories standards, exhibited a specificity of 100% and sensitivity of 92.3%. Our data show that CHO cells are a suitable host for the production of larger quantities of recombinant SARS-CoV-2 trimer which can be used as antigen for mass serological testing.
has issue date	2020-08-13(xsd:dateTime)
bibo:doi	10.1101/2020.08.07.20169441
has license	medrxiv
sha1sum (hex)	5f21f8071235a96df53e3c212bbb89171b08655d
schema:url	https://doi.org/10.1101/2020.08.07.20169441
resource representing a document's title	Production of Trimeric SARS-CoV-2 Spike Protein by CHO Cells for Serological COVID-19 Testing
resource representing a document's body	covid:5f21f8071235a96df53e3c212bbb89171b08655d#body_text
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