About: Optimizing high-yield production of SARS-CoV-2 soluble spike trimers for serology assays

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About: Optimizing high-yield production of SARS-CoV-2 soluble spike trimers for serology assays Goto Sponge NotDistinct Permalink

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

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
title	Optimizing high-yield production of SARS-CoV-2 soluble spike trimers for serology assays
Creator	Esposito, Dominic Frank, Peter Gillette, William Taylor, Troy Denson, John-Paul Drew, Matthew Gulten, Gulcin Hong, Min Mehalko, Jennifer Messing, Simon Sadtler, Kaitlyn Snead, Kelly Wall, Vanessa
source	Elsevier; Medline; PMC
abstract	The SARS-CoV-2 spike trimer is the primary antigen for several serology assays critical to determining the extent of SARS-CoV-2 exposure in the population. Until stable cell lines are developed to increase the titer of this secreted protein in mammalian cell culture, the low yield of spike protein produced from transient transfection of HEK293 cells will be a limiting factor for these assays. To improve the yield of spike protein and support the high demand for antigens in serology assays, we investigated several recombinant protein expression variables by altering the incubation temperature, harvest time, chromatography strategy, and final protein manipulation. Through this investigation, we developed a simplified and robust purification strategy that consistently yields 5 mg of protein per liter of expression culture for two commonly used forms of the SARS-CoV-2 spike protein. We show that these proteins form well-behaved stable trimers and are consistently functional in serology assays across multiple protein production lots.
has issue date	2020-06-04(xsd:dateTime)
bibo:doi	10.1016/j.pep.2020.105686
bibo:pmid	32504802
has license	no-cc
sha1sum (hex)	bd36068997ef748710fc7511a8ceb8f5e4fd15e6
schema:url	https://doi.org/10.1016/j.pep.2020.105686
resource representing a document's title	Optimizing high-yield production of SARS-CoV-2 soluble spike trimers for serology assays
has PubMed Central identifier	PMC7271859
has PubMed identifier	32504802
schema:publication	Protein Expr Purif
resource representing a document's body	covid:bd36068997ef748710fc7511a8ceb8f5e4fd15e6#body_text
is schema:about of	named entity 'trimers' named entity 'production' named entity 'serology' named entity 'assays' named entity 'assays' named entity 'PURIFICATION' named entity 'GREATER THAN' named entity 'MODIFICATION' named entity 'HKU1' named entity 'coronaviruses' named entity 'extracellular' named entity 'SARS-CoV' named entity 'transiently transfected' named entity '5.7' named entity 'supernatants' named entity 'ion chromatography' named entity 'recombinant protein' named entity 'secretory pathway' named entity 'serology' named entity 'transfection' named entity 'Mt. Sinai' named entity 'supernatants' named entity 'Coomassie staining' named entity 'expression' named entity 'greater' named entity 'serology' named entity 'spike protein' named entity 'supernatants' named entity 'spike proteins' named entity 'protein' named entity 'transiently transfected' named entity 'Absorbance' named entity 'secretion process' named entity 'biophysical' named entity 'spike proteins' named entity 'cell culture' named entity 'SEC' named entity 'GE Healthcare' named entity '37°C' named entity 'cloned' named entity 'spike proteins' named entity 'recombinant' named entity 'staining' named entity 'TFF' named entity 'immune response' named entity 'Mt. Sinai' named entity 'titer' named entity 'high affinity' named entity 'transfection' named entity 'VRC' named entity 'soluble' named entity 'soluble' named entity 'serology' named entity 'mg/l' named entity 'high-yield' named entity 'serology' named entity 'high-yield' named entity 'SARS' named entity 'structural studies' named entity 'transfection' named entity 'protein' named entity 'spike proteins' named entity 'protein' named entity 'Thermo Fisher Scientific' named entity 'spike proteins' named entity 'liquid nitrogen' named entity 'glycosylation' named entity 'infection' named entity 'antigens'

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