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10.1002/bit.27615 http://scihub22266oqcxt.onion/10.1002/bit.27615 33128388!ä!33128388 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Biotechnol+Bioeng 2021 ; 118 (2): 1013-1021 Nephropedia Template TP {{tp|p=33128388|t=2021. Production of trimeric SARS-CoV-2 spike protein by CHO cells for serological COVID-19 testing |pdf=|usr=}}{{33128388}} gab.com Text Production of trimeric SARS-CoV-2 spike protein by CHO cells for serological COVID-19 testing JO: Biotechnol Bioeng http://www.kidney.de/pget.php?&tw=1&pmid=33128388 #FOAvip We describe scalable and cost-efficient production of full length, His-tagged severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike glycoprotein trimer by Chinese hamster ovary (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 human embryonic kidney (HEK) and CHO cells mediated by polyethyleneimine was increased significantly (up to 10.9-fold) by a reduction in culture temperature to 32 degrees C 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 enzyme-linked immunosorbent assay format to detect immunoglobulin G antibodies against SARS-CoV-2 in sera from patient cohorts previously tested for viral infection by polymerase chain reaction, including those who had displayed coronavirus disease 2019 (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. Twit Text FOAVip Production of trimeric SARS-CoV-2 spike protein by CHO cells for serological COVID-19 testing ????Biotechnol Bioeng http://www.kidney.de/pget.php?&tw=1&pmid=33128388 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=33128388 #FOAvip English Wikipedia <ref>{{Cite pmid|33128388}}</ref> Production of trimeric SARS-CoV-2 spike protein by CHO cells for serological COVID-19 testing #MMPMID33128388 Johari YB; Jaffe SRP; Scarrott JM; Johnson AO; Mozzanino T; Pohle TH; Maisuria S; Bhayat-Cammack A; Lambiase G; Brown AJ; Tee KL; Jackson PJ; Wong TS; Dickman MJ; Sargur RB; James DC Biotechnol Bioeng 2021[Feb]; 118 (2): 1013-1021 PMID33128388show ga We describe scalable and cost-efficient production of full length, His-tagged severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike glycoprotein trimer by Chinese hamster ovary (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 human embryonic kidney (HEK) and CHO cells mediated by polyethyleneimine was increased significantly (up to 10.9-fold) by a reduction in culture temperature to 32 degrees C 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 enzyme-linked immunosorbent assay format to detect immunoglobulin G antibodies against SARS-CoV-2 in sera from patient cohorts previously tested for viral infection by polymerase chain reaction, including those who had displayed coronavirus disease 2019 (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. |Animals[MESH] |CHO Cells[MESH] |COVID-19 Testing/*methods[MESH] |COVID-19/*diagnosis/virology[MESH] |Cricetinae[MESH] |Cricetulus[MESH] |Humans[MESH] |Recombinant Proteins/biosynthesis[MESH] |SARS-CoV-2/*metabolism[MESH] |Serologic Tests/methods[MESH]Production of trimeric SARS-CoV-2 spike protein by CHO cells for serol(epmc).pdf DeepDyve Pubget Overpricing