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10.1016/j.pep.2020.105686 http://scihub22266oqcxt.onion/10.1016/j.pep.2020.105686 32504802!7271859!32504802     free     free     free Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Protein+Expr+Purif 2020 ; 174 (ä): 105686 Nephropedia Template TP {{tp|p=32504802|t=2020. Optimizing high-yield production of SARS-CoV-2 soluble spike trimers for serology assays |pdf=|usr=}}{{32504802}} gab.com Text Optimizing high-yield production of SARS-CoV-2 soluble spike trimers for serology assays JO: Protein Expr Purif http://www.kidney.de/pget.php?&tw=1&pmid=32504802 #FOAvip 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. Twit Text FOAVip Optimizing high-yield production of SARS-CoV-2 soluble spike trimers for serology assays ????Protein Expr Purif http://www.kidney.de/pget.php?&tw=1&pmid=32504802 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=32504802 #FOAvip English Wikipedia <ref>{{Cite pmid|32504802}}</ref> Optimizing high-yield production of SARS-CoV-2 soluble spike trimers for serology assays #MMPMID32504802 Esposito D; Mehalko J; Drew M; Snead K; Wall V; Taylor T; Frank P; Denson JP; Hong M; Gulten G; Sadtler K; Messing S; Gillette W Protein Expr Purif 2020[Oct]; 174 (ä): 105686 PMID32504802show ga 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. |Betacoronavirus/genetics/*metabolism[MESH] |COVID-19[MESH] |COVID-19 Testing[MESH] |Clinical Laboratory Techniques[MESH] |Coronavirus Infections/blood/diagnosis/virology[MESH] |Gene Expression[MESH] |HEK293 Cells[MESH] |Humans[MESH] |Pandemics[MESH] |Pneumonia, Viral/blood/diagnosis/virology[MESH] |Protein Stability[MESH] |Recombinant Proteins/chemistry/genetics/metabolism[MESH] |SARS-CoV-2[MESH] |Spike Glycoprotein, Coronavirus/chemistry/*genetics/*isolation & purification/metabolism[MESH]Optimizing high-yield production of SARS-CoV-2 soluble spike trimers f(epmc).pdf DeepDyve Pubget Overpricing